Azacyclic derivative, preparation method therefor and use thereof

Abstract

The present invention relates to an azacyclic derivative, a preparation method therefor and a use of a pharmaceutical composition containing the derivative in medicine. Specifically, the present invention relates to a cyclic derivative represented by general formula (I), a preparation method therefor, a pharmaceutically acceptable salt thereof, and a use thereof as a therapeutic agent, especially as a WRN inhibitor. The definition of each substituent in general formula (I) is the same as that in the description.

Description

Azepino derivatives, processes for their preparation and uses thereof

[0001] Cross-reference to related applications

[0002] This application claims priority to the following patent applications: 1) a patent application entitled “Azepino derivatives, processes for their preparation and uses thereof” filed on August 22, 2024 with the China National Intellectual Property Office, application number CN202411156399.2; 2) a patent application entitled “Azepino derivatives, processes for their preparation and uses thereof” filed on January 27, 2025 with the China National Intellectual Property Office, application number CN202510126777.0; and 3) a patent application entitled “Azepino derivatives, processes for their preparation and uses thereof” filed on April 9, 2025 with the China National Intellectual Property Office, application number CN202510446767.5, the contents of all of the above patent applications are incorporated herein by reference in their entirety. TECHNICAL FIELD

[0003] The present application relates to azepino derivatives, processes for their preparation and pharmaceutical compositions containing them and their use as therapeutic agents, in particular as WRN inhibitors. BACKGROUND

[0004] WRN in humans is composed of 1432 amino acid residues, which contains five important components from N-terminal to C-terminal, including exonuclease domain, ATPase domain, RecQ C-terminal domain, helicase / ribonuclease D C-terminal domain and nuclear localization signal. Among the five human RecQ helicases, WRN is the only one with 3'→5' exonuclease activity, which is achieved by the specific activation of its N-terminal exonuclease domain by Ku70 / 80 complex bound to the DNA terminus. The ATPase domain is the largest and most conserved component in the RecQ helicase family, which acts as an ATP-dependent DNA translocation module by binding and hydrolyzing ATP. The RecQ C-terminal domain is the main site of DNA binding and catalyzes the unwinding of DNA double strands. Therefore, the ATPase domain and the RecQ C-terminal domain together constitute the core of the WRN helicase. WRN is a DNA helicase with multiple enzyme activities that can bind to DNA and other proteins. This makes the enzyme play an important role in maintaining the integrity and stability of the genome, including participating in DNA damage repair, replication and transcription, and maintaining telomere and heterochromatin stability.

[0005] Synthetic lethality refers to the phenomenon that two non-lethal genes are simultaneously inhibited (inhibited forms include gene defects such as gene mutation, gene silencing, and / or molecular perturbations such as gene expression knock-out, drug inhibition) to cause cell death. Using this mechanism, a specific mutation in cancer is found, and its "synthetic lethal partner" is found and inhibited, thereby specifically killing cancer cells with the mutation.

[0006] Studies have shown that WRN is a "synthetic lethal partner" of high microsatellite instability (MSI-H), a type of genomic damage. High microsatellite instability (MSI-H) is a hyper-variable state caused by frequent insertion and / or deletion mutations in nucleotide repeat regions due to defects in DNA mismatch repair (MMR), which is commonly seen in endometrial cancer (31%), colorectal cancer (25%), and gastric cancer (19%) and other cancers. In MSI-H cancer cells, thymine / adenine dinucleotide (TA) repeat sequences are highly unstable and undergo large-scale amplification, forming non-canonical right-handed double helix (non-B) DNA secondary structures (such as cruciform and G-quadruplex), which require WRN-specific unwinding to complete replication. In the absence of WRN, these DNA secondary structures are cut by the MUS81-EME1-SLX4 endonuclease complex, leading to extensive DNA end resection, depletion of replication protein A (RPA), chromosomal fragmentation, and cell death. In addition, in tumor models with MMR defects, the absence of WRN leads to the activation of multiple DNA damage signaling markers, inducing cell cycle arrest and apoptosis, and thereby inhibiting the proliferation of tumor cells. Recent studies have shown that WRN small molecule inhibitors can specifically cause tumor regression in MSI-H tumor models, but have no effect in microsatellite stable (MSS) tumor models. Therefore, small molecule chemicals that can inhibit WRN helicase activity are expected to become a new method for effectively treating MSI-H cancer.

[0007] There is no drug on the market for WRN target inhibitors, and there is still a lack of effective treatment drugs for tumor diseases, which is a serious threat to human health, and there is still a huge unmet clinical need. SUMMARY

[0008] To solve the above technical problems, the present application provides a compound represented by general formula (I) or a stereoisomer, tautomer, deuterated product or pharmaceutically acceptable salt thereof:

[0009] wherein:

[0010] is selected from a single or double bond as necessary to render each atom thereof a normal valence;

[0011] bond denotes may exist as (Z)- or (E)-stereoisomers, wherein

[0012] ring A is a 5- to 12-membered heterocyclyl or a 5- to 6-membered heteroaryl, wherein said heterocyclyl or heteroaryl contains at least one nitrogen atom;

[0013] X is selected from CR a and N;

[0014] Z is selected from CR c and N;

[0015] or, is absent (i.e. is )

[0016] Y is selected from CR b , N, NR d , S and O;

[0017] or, Y and Z together form a C 3-7 cycloalkyl, 3- to 7-membered heterocyclyl or 5- to 6-membered heteroaryl, said C 3-7 cycloalkyl, 3- to 7-membered heterocyclyl or 5- to 6-membered heteroaryl is optionally further substituted by one or more substituents selected from a deuterium atom, hydroxy, halogen, nitro, cyano, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR 5 , =0, -C(=0)R 5 , -C(=0)OR 5 , -OC(=0)R 5 , -NR 6 R 7 , -C(=0)NR 6 R 7 , -S(=0)2NR 6 R 7 , -N(R 6 )C(=0)R 7 and -N(R 6 )C(=0)OR 7 ;

[0018] R a , R b , R c , Rd Each is independently selected from hydrogen atoms, halogens, cyano groups, and C atoms. 1-6 Alkyl and C 3-6 cycloalkyl, wherein the C 1-6 Alkyl or C 3-6 The cycloalkyl group may optionally be further selected from one or more halogens, hydroxyl groups, cyano groups, and C. 1-6 Substituents of alkoxy groups;

[0019] R 1 Whether the groups are the same or different, they are each independently selected from deuterium, hydroxyl, halogen, nitro, cyano, and C atoms. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, 5-10 heterocyclic, C 6-10 Aryl, 5-6 quinone heteroaryl, -SF5, -OR 5 -OC(=O)R 5 -C(=O)R 5 -C(=O)OR 5 -N(R) 6 )C(=O)R 7 -N(R) 6 )C(=O)OR 7 -NR 6 R 7 -C(=O)NR 6 R 7 -S (=O) r NR 6 R 7 Or -S (=O) r R 5 The C mentioned therein 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3- 8-cycloalkyl, 5-10 heterocyclic, C 6-10 aryl, 5-6 heteroaryl, optionally further selected by one or more groups selected from hydroxyl, halogen, nitro, cyano, alkyl, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, -OR 5 =O, -C(=O)R 5 -C(=O)OR 5 -OC(=O)R 5 -NR 6 R 7 -C(=O)NR 6 R 7 -S(=O)2NR 6 R 7 -N(R)6 )C(=O)R 7 and -N(R 6 )C(=O)OR 7 substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, nitro, cyano, C

[0020] and / or, two R 1 form together with the carbon atom to which they are attached a -C(=O);

[0021] R 2 is selected from the group consisting of hydrogen atom, deuterium atom, hydroxyl, halogen, nitro, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, 5-10 membered heterocyclyl, C 6-10 aryl, 5-6 membered heteroaryl, -SF5, -OR 5 , -OC(=O)R 5 , -C(=O)R 5 , -C(=O)OR 5 , -N(R 6 )C(=O)R 7 , -N(R 6 )C(=O)OR 7 , -NR 6 R 7 , -C(=O)NR 6 R 7 , -S(=O) r NR 6 R 7 and -S(=O) r R 5 , wherein said C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, 5-10 membered heterocyclyl, C 6-10 aryl, 5-6 membered heteroaryl are optionally further substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR 5 , =O, -C(=O)R 5 , -C(=O)OR 5 , -OC(=O)R 5 , -NR 6 R 7 , -C(=O)NR 6 R 7 , -S(=O)2NR 6 R 7 , -N(R 6)C(=O)R 7 and -N(R 6 )C(=O)OR 7 substituted by one or more substituents selected from the group consisting of deuterium atom, hydroxy, halogen, nitro, cyano, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR

[0022] or, R 2 and Y, R 2 and X are each independently, together with the atom to which they are attached, a C 3-7 cycloalkyl, 3-7 membered heterocyclyl, C 6-10 aryl or 5-6 membered heteroaryl, said C 3-7 cycloalkyl, 3-7 membered heterocyclyl, C 6-10 aryl or 5-6 membered heteroaryl is optionally further substituted by one or more substituents selected from the group consisting of deuterium atom, hydroxy, halogen, nitro, cyano, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR 5 , =O, -C(=O)R 5 , -C(=O)OR 5 , -OC(=O)R 5 , -NR 6 R 7 , -C(=O)NR 6 R 7 , -S(=O)2NR 6 R 7 , -N(R 6 )C(=O)R 7 and -N(R 6 )C(=O)OR 7 substituted by one or more substituents selected from the group consisting of deuterium atom, hydroxy, halogen, nitro, cyano, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR

[0023] R g is selected from the group consisting of hydrogen atom and deuterium atom, preferably hydrogen atom;

[0024] R f are each independently selected from the group consisting of hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl and 5-10 membered heterocyclyl; wherein C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl or 5-10 membered heterocyclyl is optionally further substituted by one or more substituents selected from the group consisting of halogen, hydroxy, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy and C 1-6 haloalkoxy;

[0025] R 3a , R 3b are each independently selected from the group consisting of hydrogen atom and C 1-6 alkyl;

[0026] R 4 is selected from C 1-6 alkyl, C 3-8 cycloalkyl and 5-10 membered heterocyclyl; wherein C 1-6 alkyl, C 3-8 cycloalkyl or 5-10 membered heterocyclyl is optionally further substituted with one or more substituents selected from the group consisting of halogen, hydroxy, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1- 6alkoxy and C 1-6 haloalkoxy;

[0027] R 5 each independently is selected from the group consisting of alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further substituted with one or more substituents selected from the group consisting of a deuterium atom, hydroxy, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -C(=0)R 8 , -C(=0)OR 8 , -OC(=0)R 8 , -NR 9 R 10 , -C(=0)NR 9 R 10 , -S(=0)2NR 9 R 10 and -N(R 9 )C(=0)R 10 ;

[0028] R 6 and R 7 each independently is selected from the group consisting of a hydrogen atom, hydroxy, alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further substituted with one or more substituents selected from the group consisting of hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -C(=0)R 8 , -C(=0)OR 8 , -OC(=0)R 8 , -NR 9 R 10 , -C(=0)NR 9 R 10 , -S(=0)2NR 9 R 10 and -N(R 9 )C(=0)R 10 ;

[0029] or, R 6 and R 7 together with the atom to which they are attached form a 4-8 membered heterocyclyl group containing one or more N, O or S(=O) r wherein said 4-8 membered heterocyclyl group is optionally further substituted by one or more substituents selected from the group consisting of hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -C(=O)R 8 , -C(=O)OR 8 , -OC(=O)R 8 , -NR 9 R 10 , -C(=O)NR 9 R 10 , -S(=O)2NR 9 R 10 and -N(R 9 )C(=O)R 10 ;

[0030] R 8 , R 9 and R 10 are each independently selected from the group consisting of hydrogen atom, alkyl, amino, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further substituted by one or more substituents selected from the group consisting of hydroxy, halogen, nitro, amino, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxy and carboxylate;

[0031] n is 0, 1, 2, 3 or 4; and

[0032] each r is independently 0, 1 or 2.

[0033] In a preferred embodiment of the present application, a compound of general formula (I) or a stereoisomer, a tautomer, a deuterated analogue or a pharmaceutically acceptable salt thereof, wherein,

[0034] X is selected from CR a and N;

[0035] Y is selected from CR b and N;

[0036] Z is selected from CR c and N;

[0037] and at least one of X, Y, Z is N and X, Y, Z are not simultaneously N;

[0038] or, Y and Z together form a C 3-7 cycloalkyl, 3-7 membered heterocyclyl or 5-6 membered heteroaryl, said C 3-7cycloalkyl, 3- to 7-membered heterocyclyl, or 5- to 6-membered heteroaryl is optionally further substituted with one or more substituents selected from the group consisting of a deuterium atom, a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, a haloalkyl group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, -OR 5 , =0, -C(=0)R 5 , -C(=0)OR 5 , -OC(=0)R 5 , -NR 6 R 7 , -C(=0)NR 6 R 7 , -S(=0)2NR 6 R 7 , -N(R 6 )C(=0)R 7 , and -N(R 6 )C(=0)OR 7 ;

[0039] R a , R b , R c , R 5 , R 6 , and R 7 are as described in general formula (I).

[0040] According to an embodiment of the present application, a compound of general formula (I) or a stereoisomer, a tautomer, a deuterated analog, or a pharmaceutically acceptable salt thereof, wherein Z is selected from CH.

[0041] According to an embodiment of the present application, a compound of general formula (I) or a stereoisomer, a tautomer, a deuterated analog, or a pharmaceutically acceptable salt thereof, is a compound of general formula (II) or a stereoisomer, a tautomer, a deuterated analog, or a pharmaceutically acceptable salt thereof:

[0042] wherein:

[0043] X, Y are each independently selected from CH and N; and at least one of X, Y is N;

[0044] as required to result in each atom having a normal valency;

[0045] R 3a , R 3b are each independently selected from a hydrogen atom;

[0046] ring A, R 1 , R 2 , R g , R f , R 4and n are as defined in claim 1.

[0047] In a preferred embodiment of the present application, a compound according to formula (I) or (II), wherein Z is CH, X and Y are CH, or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof, is a compound according to formula (III), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof:

[0048] wherein:

[0049] X, Y are each independently selected from CH and N; and at least one of X, Y is N;

[0050] is selected from a single and a double bond, as required, so that each atom thereof has a normal valency;

[0051] R 3a , R 3b are each independently selected from a hydrogen atom;

[0052] ring A, R 1 , R 2 , R g , R f , R 4 and n are as defined in formula (I).

[0053] In a preferred embodiment of the present application, a compound according to formula (I), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof, is a compound according to formula (III), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof:

[0054] In a preferred embodiment of the present application, a compound according to formula (I), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof, is a compound according to formula (IV), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof:

[0055] wherein:

[0056] is selected from a single and a double bond, as required, so that each atom thereof has a normal valency;

[0057] R 3a , R 3b are each independently selected from a hydrogen atom;

[0058] ring A, R 1 , R 2 , R g , R f , R 4 and n are as defined in formula (I).

[0059] In a preferred embodiment of the present application, a compound according to formula (I) or (IV), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof, is a compound according to formula (IV-a), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof:

[0060] wherein: R 3a , R 3b , ring A, R 1 , R 2 , R g , R f , R 4 and n are as defined in formula (IV).

[0061] In a preferred embodiment of the present application, a compound according to formula (I), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof, is a compound according to formula (I) wherein Z is selected from N.

[0062] In a preferred embodiment of the present application, a compound according to formula (I), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof, is a compound according to formula (V), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof:

[0063] wherein X, Y are each independently selected from CH and N; and X, Y are not simultaneously N;

[0064] as required to result in each atom having a normal valency;

[0065] R 3a , R 3b are each independently selected from a hydrogen atom;

[0066] ring A, R 1 , R 2 , R g , R f , R 4 or n are as defined in formula (I).

[0067] In a preferred embodiment of the present application, a compound according to formula (I) or (V), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof, is a compound according to formula (V-a), or a stereoisomer, a tautomer, a deuterated analogue, or a pharmaceutically acceptable salt thereof:

[0068] wherein X, Y, R 3a , R 3b , ring A, R1 2 g f 4 and n are defined as in general formula (V).

[0069] According to a preferred embodiment of the present application, a compound according to general formula (I) or a stereoisomer, a tautomer, a deuterated analogue or a pharmaceutically acceptable salt thereof, is a compound according to general formula (II) or a stereoisomer, a tautomer, a deuterated analogue or a pharmaceutically acceptable salt thereof: is absent.

[0070] Accordingly, according to a preferred embodiment of the present application, a compound according to general formula (I) or a stereoisomer, a tautomer, a deuterated analogue or a pharmaceutically acceptable salt thereof, is a compound according to general formula (VI) or a stereoisomer, a tautomer, a deuterated analogue or a pharmaceutically acceptable salt thereof:

[0071] wherein

[0072] is selected from a single and a double bond, as required, to give each atom thereof a normal valency;

[0073] Y is selected from NR d , S and O;

[0074] R d is selected from a hydrogen atom and C 1-6 alkyl, wherein said C 1-6 alkyl is optionally further substituted by one or more substituents selected from halogen, hydroxyl, cyano and C 1-6 alkoxy;

[0075] R 2 is selected from hydroxyl, halogen, cyano, C 1-6 alkyl, C 3-8 cycloalkyl and C 1-6 alkoxy, wherein said C 1-6 alkyl, C 3-8 cycloalkyl or C 1-6 alkoxy is optionally further substituted by one or more halogen, C 1-6 alkyl or C 3-8 cycloalkyl.

[0076] X is selected from CH and N;

[0077] or, R 2 and X, together with the atom to which they are attached, form a C 6-10 aryl or 5- to 6-membered heteroaryl, wherein said C 6-10 aryl or 5- to 6-membered heteroaryl is optionally further substituted by one or more substituents selected from a deuterium atom, hydroxyl, halogen, nitro, cyano, C 1-6 alkyl, C​​​​1-6 Haloalkyl, C 3-8 cycloalkyl and C 1-6 Substituents of alkoxy groups;

[0078] Or, R 2 Y and the atoms it is attached to together form C 6-10 Aryl or 5-6 heteroaryl, wherein the C 6-10 aryl or 5-6 heteroaryl groups may be further selected from one or more deuterium atoms, hydroxyl groups, halogens, nitro groups, cyano groups, C6 groups, etc. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 cycloalkyl and C 1-6 Substituents of alkoxy groups;

[0079] Rings A and R 1 R 2 R g R f R 4 R 3a R 3b Or n is defined as in general formula (I).

[0080] A preferred embodiment of the present invention is a compound of general formula (I) or (VI) or a stereoisomer, tautomer, deuterated derivative or pharmaceutically acceptable salt thereof, wherein the compound is of general formula (VI-a) or a stereoisomer, tautomer, deuterated derivative or pharmaceutically acceptable salt thereof:

[0081] in,

[0082] R 2 X, Y, ring A, R 1 R 2 R g R f R 4 R 3a R 3b The definitions of n are as described in general formula (VI).

[0083] A preferred embodiment of the present invention is a compound of general formula (I) or (VI) or a stereoisomer, tautomer, deuterated derivative or pharmaceutically acceptable salt thereof, wherein the compound is of general formula (VI-1) or a stereoisomer, tautomer, deuterated derivative or pharmaceutically acceptable salt thereof:

[0084] in, The atoms are selected from single and double bonds as needed to ensure that each atom is in the normal valence state;

[0085] X1, X2, X3are each independently selected from CR e and N, and X1, X2, X3are not simultaneously N;

[0086] Y is selected from NR d , S and O;

[0087] R d is selected from a hydrogen atom and C 1-6 alkyl;

[0088] R e are each independently selected from a hydrogen atom, a deuterium atom, a hydroxyl group, a halogen, a nitro group, a cyano group, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and C 1-6 alkoxy;

[0089] Ring A, R 1 , R g , R f , R 4 , R 3a , R 3b and n are as described in general formula (I).

[0090] In a preferred embodiment of the present application, a compound according to general formula (I), (VI) or (VI-1) or a stereoisomer, a tautomer, a deuterated compound thereof, or a pharmaceutically acceptable salt thereof, is a compound according to general formula (VI-1a) or a stereoisomer, a tautomer, a deuterated compound thereof, or a pharmaceutically acceptable salt thereof:

[0091] wherein, X1, X2, X3, Y, Ring A, R 1 , R g , R f , R 4 , R 3a , R 3b and n are as described in general formula (VI-1).

[0092] In a preferred embodiment of the present application, a compound according to general formula (I) or (VI) or a stereoisomer, a tautomer, a deuterated compound thereof, or a pharmaceutically acceptable salt thereof, is a compound according to general formula (VI-2) or a stereoisomer, a tautomer, a deuterated compound thereof, or a pharmaceutically acceptable salt thereof:

[0093] wherein, X4, X5, X6are each independently selected from CR e and N, and X4, X5, X6are not simultaneously N;

[0094] X is selected from CH or N;

[0095] R e each independently is selected from the group consisting of a hydrogen atom, a deuterium atom, a hydroxyl group, a halogen, a nitro group, a cyano group, a C 1-6 alkyl group, a C 1-6 haloalkyl group, a C 3-8 cycloalkyl group, or a C 1-6 alkoxy group;

[0096] ring A, R 1 , R g , R f , R 4 , R 3a , R 3b and n are defined as described in general formula (I).

[0097] In a preferred embodiment of the present application, a compound according to general formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a), or a stereoisomer, a tautomer, a deuterated analog, or a pharmaceutically acceptable salt thereof, is a compound according to general formula (VI-2a), or a stereoisomer, a tautomer, a deuterated analog, or a pharmaceutically acceptable salt thereof:

[0098] wherein, X4, X5, X6, X, ring A, R 1 , R g , R f , R 4 , R 3a , R 3b or n are defined as described in general formula (VI-2).

[0099] In a preferred embodiment of the present application, a compound according to general formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a), or a stereoisomer, a tautomer, a deuterated analog, or a pharmaceutically acceptable salt thereof, is a compound according to general formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a), wherein is selected from the group consisting of:

[0100] In a preferred embodiment of the present application, a compound according to general formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a), or a stereoisomer, a tautomer, a deuterated analog, or a pharmaceutically acceptable salt thereof, is a compound according to general formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a), wherein R 1 are the same or different, each independently selected from the group consisting of a C 6-10 aryl group and a C 3-8 cycloalkyl group, wherein said C 6-10 aryl group and C 3-8Cycloalkyl is optionally further substituted by one or more substituents selected from the group consisting of halogen, cyano, amino, C 1-6 Alkyl and C 1-6 Alkoxy, and / or two R

[0101] and / or, two R 1 form together with the carbon atom to which they are attached a -C(=0).

[0102] In a preferred embodiment of the present application, a compound according to formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a) or a stereoisomer, a tautomer, a deuterated derivative or a pharmaceutically acceptable salt thereof, wherein R 1 are each independently selected from the group consisting of C 6-10 Aryl, more preferably phenyl,

[0103] and / or, two R 1 form together with the carbon atom to which they are attached a -C(=0).

[0104] In a preferred embodiment of the present application, a compound according to formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a) or a stereoisomer, a tautomer, a deuterated derivative or a pharmaceutically acceptable salt thereof, wherein is selected from the group consisting of:

[0105] R 1a are each independently selected from the group consisting of C 6-10 Aryl and C 3-8 Cycloalkyl, wherein said C 6-10 Aryl and C 3-8 Cycloalkyl is optionally further substituted by one or more substituents selected from the group consisting of halogen, cyano, amino, C 1-6 Alkyl and C 1-6 Alkoxy, and / or two R

[0106] Preferably, R 1a is selected from the group consisting of phenyl, More preferably, R 1a is phenyl.

[0107] In a preferred embodiment of the present application, a compound according to formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a) or a stereoisomer, a tautomer, a deuterated derivative or a pharmaceutically acceptable salt thereof, wherein R g is a hydrogen atom; R f is selected from the group consisting of C 1-6 alkyl and C 3-8 cycloalkyl, wherein said C 1-6 alkyl is optionally further substituted by one C 1-6 alkoxy, wherein said C 3-8 cycloalkyl is optionally further substituted by one or more halogen, said C 3-8 cycloalkyl is preferably cyclopropyl or cyclobutyl.

[0108] In a preferred embodiment of the present application, a compound according to formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a) or a stereoisomer, a tautomer, a deuterated derivative or a pharmaceutically acceptable salt thereof, wherein R 4 is selected from the group consisting of C 1-6 alkyl, preferably methyl.

[0109] In a preferred embodiment of the present application, a compound according to formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a) or a stereoisomer, a tautomer, a deuterated derivative or a pharmaceutically acceptable salt thereof, wherein R 2 is selected from the group consisting of a hydrogen atom, hydroxy, halogen, cyano, -NR 6 R 7 , C 1-6 alkyl, C 3-8 cycloalkyl and C 1-6 alkoxy, wherein said C 1- 6alkyl, C 3-8 cycloalkyl and C 1-6 alkoxy is optionally further substituted by one or more halogen, C 1-6 alkyl or C 3-8 cycloalkyl;

[0110] R 6 , R 7 are each independently selected from the group consisting of a hydrogen atom and C 1-6 alkyl.

[0111] A preferred embodiment of the present invention is a general formula (I), (II), (III), (IV), (IV-a), (V), (Va), (VI), (VI-a) wherein R 2 Selected from hydrogen atom, hydroxyl group, fluorine, chlorine, bromine, cyano, methyl, tert-butyl, methoxy, trifluoromethyl, difluoromethyl, trifluoromethoxy, dimethylamino,

[0112] In a preferred embodiment of the present invention, the compounds of the general formula are selected from:

[0113] Or its stereoisomers, tautomers, or medicinal salts. Note: In case of discrepancy between the drawn structure and the given name of the structure, the drawn structure shall prevail.

[0114] Furthermore, the present invention provides a pharmaceutical composition comprising a compound of general formula (I), (II), (III), (IV), (IV-a), (V), (Va), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0115] The present invention provides the use of a compound of general formula (I), (II), (III), (IV), (IV-a), (V), (Va), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a WRN inhibitor.

[0116] The present application also provides a use of a compound of Formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a), or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the manufacture of a medicament for treating a WRN-mediated disease, wherein the WRN-mediated disease is preferably a microsatellite instability-high (MSI-H) cancer; and wherein the WRN-mediated disease is selected from the group consisting of colorectal cancer, gastric cancer, endometrial cancer, rectal adenocarcinoma, adrenocortical carcinoma, uterine sarcoma, cervical cancer, nephroblastoma, mesothelioma, esophageal cancer, breast cancer, renal clear cell carcinoma, ovarian serous cystadenocarcinoma, cholangiocarcinoma, thymoma, liver cancer, head and neck squamous cell carcinoma, sarcoma, cutaneous melanoma, lung squamous cell carcinoma, prostate cancer, lung adenocarcinoma, bladder transitional cell carcinoma, pediatric neuroblastoma, chronic lymphocytic leukemia and glioma, preferably colorectal cancer, gastric cancer or endometrial cancer.

[0117] The present application further provides a use of a compound of Formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a), or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the manufacture of a medicament for treating a microsatellite instability-high (MSI-H) cancer.

[0118] The present application provides a use of a compound of Formula (I), (II), (III), (IV), (IV-a), (V), (V-a), (VI), (VI-a), (VI-1), (VI-1a), (VI-2) or (VI-2a), or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the manufacture of a medicament for treating colorectal cancer, gastric cancer, endometrial cancer, rectal adenocarcinoma, adrenocortical carcinoma, uterine sarcoma, cervical cancer, nephroblastoma, mesothelioma, esophageal cancer, breast cancer, renal clear cell carcinoma, ovarian serous cystadenocarcinoma, cholangiocarcinoma, thymoma, liver cancer, head and neck squamous cell carcinoma, sarcoma, cutaneous melanoma, lung squamous cell carcinoma, prostate cancer, lung adenocarcinoma, bladder transitional cell carcinoma, pediatric neuroblastoma, chronic lymphocytic leukemia or glioma, preferably in the manufacture of a medicament for treating colorectal cancer, gastric cancer or endometrial cancer.

[0119] Accordingly, this application also provides a method for treating or preventing WRN-mediated diseases, comprising administering to a subject in need a compound of general formula (I), (II), (III), (IV), (IV-a), (V), (Va), (VI), (VI-a), (VI-1), (VI-1a), (VI-2), or (VI-2a) of this application, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in this application. The WRN-mediated disease is preferably a highly microsatellite unstable cancer. Accordingly, this application also provides a method for treating or preventing highly microsatellite unstable cancer, comprising administering to a subject in need a compound of general formula (I), (II), (III), (IV), (IV-a), (V), (Va), (VI), (VI-a), (VI-1), (VI-1a), (VI-2), or (VI-2a) of this application, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in this application. The diseases mediated by WRN can be selected from colorectal cancer, gastric cancer, endometrial cancer, rectal adenocarcinoma, adrenocortical carcinoma, uterine sarcoma, cervical cancer, nephroblastoma, mesothelioma, esophageal cancer, breast cancer, clear cell renal cell carcinoma, ovarian serous cystadenocarcinoma, bile duct carcinoma, thymoma, liver cancer, head and neck squamous cell carcinoma, sarcoma, skin melanoma, lung squamous cell carcinoma, prostate cancer, lung adenocarcinoma, bladder transitional cell carcinoma, pediatric neuroblastoma, chronic lymphocytic leukemia, and glioma, and more preferably colorectal cancer, gastric cancer, or endometrial cancer.

[0120] Detailed description of the invention

[0121] Unless otherwise stated, some terms used in this specification and claims are defined as follows:

[0122] When "alkyl" is used as a group or part of a group, it refers to a group consisting of C1-C2. 20 Straight-chain or branched aliphatic hydrocarbon groups. Preferably C1-C. 10 Alkyl groups, more preferably C1-C6 or C1-C4 alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. Alkyl groups may be substituted or unsubstituted.

[0123] "Alkenyl" refers to an alkyl group as defined above, consisting of at least two carbon atoms and at least one carbon-carbon double bond. Representative examples include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl. C2-C4 alkenyl groups are preferred. Alkenyl groups may be optionally substituted or unsubstituted.

[0124] "Alkyne group" refers to an aliphatic hydrocarbon group containing a single carbon-carbon triple bond, which can be straight-chain or branched. C2-C is preferred. 10 The alkynyl group is preferred, more preferably C2-C6 alkynyl, and most preferably C2-C4 alkynyl. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl. The alkynyl group may be substituted or unsubstituted.

[0125] "Cycloalkyl" refers to a non-aromatic cyclic alkyl group in which one or more cyclic atoms are carbon atoms, and the ring contains 0, 1, or more double bonds, including monocyclic, polycyclic, fused, bridged, and spirocyclic rings, preferably having a 3- to 7-membered monocyclic ring or a 5- to 18-membered bicyclic or tricyclic ring. In this document, cycloalkyl can be C 3-12 Cycloalkyl groups, for example, cycloalkyl groups having 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms; in some embodiments, the cycloalkyl group may be C16-126. 5-12 cycloalkyl, C 3-6 cycloalkyl, C 3-7 cycloalkyl, C 3-8 cycloalkyl or C 5-6 Cycloalkyl group. In some embodiments, the cycloalkyl group is cyclopropyl.

[0126] Examples of "monocycloalkyl" include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0127] Monocyclic alkyl groups can be substituted or unsubstituted.

[0128] "Spirocycloalkyl" refers to a polycyclic group consisting of 5 to 18 quintiles, two or more cyclic structures, where the monocyclic rings share a carbon atom (called a spiro atom) with each other, containing 0, 1, or more double bonds within the rings, but without any ring having fully conjugated π electrons. Preferably, it is a 6 to 14 quintile group, more preferably a 7 to 10 quintile group. Based on the number of spiro atoms shared between the rings, spirocycloalkyl groups are classified as monospiro, bispiro, or polyspirocycloalkyl groups, preferably monospiro and bispirocycloalkyl groups, and preferably 4 / 5, 4 / 4, 4 / 6, 3 / 6, 5 / 5, or 5 / 6 quintile groups. Non-limiting examples of "spirocycloalkyl" include, but are not limited to: spiro[4.5]decyl, spiro[4.4]nonyl, spiro[3.5]nonyl, spiro[2.4]heptyl, Cycloalkyl groups can be substituted or unsubstituted.

[0129] “Fused cycloalkyl” refers to a fully carbon polycyclic group of 5 to 18 members, containing two or more cyclic structures sharing a pair of carbon atoms with each other, one or more rings can contain 0, 1, or multiple double bonds, but no ring has a fully conjugated system of pi electrons, preferably 6 to 14 members, more preferably 6 to 10 members. It can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused cycloalkyl according to the number of rings that make up the group, preferably bicyclic or tricyclic, more preferably 3 / 5, 5 / 5, or 5 / 6 bicyclic fused cycloalkyl. Non-limiting examples of “fused cycloalkyl” include, but are not limited to: bicyclo[3.1.0]hexyl, bicyclo[3.2.0]hept-1-enyl, bicyclo[3.2.0]heptyl, decahydronaphthyl, tetradecahydrophenanthryl, Fused cycloalkyl groups can be substituted or unsubstituted.

[0130] “Bridged cycloalkyl” refers to a fully carbon polycyclic group of 5 to 18 members, containing two or more cyclic structures sharing two non-directly connected carbon atoms with each other, one, two, or three rings can contain 0, 1, or multiple double bonds, but no ring has a fully conjugated system of pi electrons, preferably 6 to 14 members, more preferably 7 to 10 members. It can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl according to the number of rings that make up the group, preferably bicyclic, tricyclic, or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of “bridged cycloalkyl” include, but are not limited to: (1s,4s)-bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, (1s,5s)-bicyclo[3.3.1]nonyl, bicyclo[2.2.2]octyl, (1r,5r)-bicyclo[3.3.2]decyl, Bridged cycloalkyl groups can be substituted or unsubstituted.

[0131] “Heterocyclyl,” “heterocycloalkyl,” “heterocycle,” or “heterocyclic” are used interchangeably herein and refer to non-aromatic heterocyclic groups in which 1, 2, or 3 ring-forming atoms are selected from nitrogen, oxygen, and S(O) r (wherein r is selected from 0, 1, and 2) heteroatoms, containing 0, 1, or multiple double bonds within the ring, including monocyclic, polycyclic, fused, bridged, and spirocyclic. Heterocyclyl groups herein can contain 1, 2, or 3 atoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the heterocyclyl group contains 1, 2, or 3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the heterocyclyl group has a ring member count of 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In some embodiments, the heterocyclyl group is a 3-12 membered heterocyclyl, 3-10 membered heterocyclyl, 5-12 membered heterocyclyl, 5-10 membered heterocyclyl, 3-7 membered heterocyclyl, 4-8 membered heterocyclyl, or 5-6 membered heterocyclyl.

[0132] Heterocyclyl groups can be substituted or unsubstituted.

[0133] Examples of "monocyclic heterocyclyl groups" include, but are not limited to, morpholinyl, oxetanyl, azetidinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidinyl, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octanyl, piperazinyl, hexahydropyrimidinyl,

[0134] "Spiroheterocyclyl" refers to a polycyclic group of 5 to 18 members, two or more cyclic structures, and one atom in common between the single rings, containing 0, 1, or more double bonds within the rings, but no ring having a fully conjugated system of π electrons, wherein one or more ring atoms are selected from nitrogen, oxygen, and S(O) r heteroatoms, the remaining ring atoms being carbon. Preferably 6 to 14 membered, more preferably 7 to 10 membered. Spirocycloalkyl groups are classified as mono-, bi- or polyspirocycloalkyl groups, preferably mono- and bi-spirocycloalkyl groups, according to the number of spiro atoms in common between the rings. More preferably 3 / 6, 4 / 4, 4 / 5, 4 / 6, 5 / 5, 5 / 6, or 6 / 6 membered mono-spirocycloalkyl groups. Non-limiting examples of "spiroheterocyclyl groups" include, but are not limited to: 1,7-dioxaspiro[4.5]decanyl, 2-oxa-7-azaspiro[4.4]nonanyl, 7-oxaspiro[3.5]nonanyl, 5-oxaspiro[2.4]heptanyl,

[0135] "Fused heterocyclyl" refers to a polycyclic group of two or more cyclic structures sharing a pair of atoms in common between the rings, one or more rings can contain 0, 1, or more double bonds, but no ring having a fully conjugated system of π electrons, wherein one or more ring atoms are selected from nitrogen, oxygen, and S(O) r heteroatoms, the remaining ring atoms being carbon. Preferably 6 to 14 membered, more preferably 7 to 10 membered. Fused heterocyclyl groups are classified as bi-, tri-, tetra- or polycyclic, preferably bi- or tri-cyclic, more preferably 5 / 5 or 5 / 6 bi-cyclic fused heterocyclyl groups, according to the number of rings making up the group. Non-limiting examples of "fused heterocyclyl groups" include, but are not limited to: octahydropyrrolo[3,4-c]pyrrolyl, octahydro-1H-isoindolyl, 3-azabicyclo[3.1.0]hexanyl, octahydrobenzo[b][1,4]dioxine,

[0136] "bridged heterocyclyl" means a polycyclic group of 5 to 18 members, containing two or more cyclic structures, sharing two non-adjacent atoms with each other, one or more rings can contain 0, 1 or more double bonds, but none of the rings has a fully conjugated π-electron aromatic system, wherein one or more ring atoms are selected from nitrogen, oxygen and S(O) r ranging from 6 to 14 members, more preferably from 7 to 10 members. Depending on the number of rings, it can be a bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclyl, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged heterocyclyl" include, but are not limited to: 2-azabicyclo[2.2.1]heptyl, 2-azabicyclo[2.2.2]octyl, 2-azabicyclo[3.3.2]decyl.

[0137] "aryl" means a carbocyclic aromatic system containing one or two rings, wherein the rings can be connected together in a fused manner. The term "aryl" includes mono- or bicyclic aromatic groups such as the aromatic groups of phenyl, naphthyl, tetrahydronaphthyl. Preferably, aryl is C6-C 10 aryl, more preferably aryl is phenyl and naphthyl, most preferably naphthyl. The aryl group can be substituted or unsubstituted.

[0138] "Heteroaryl" means an aromatic 5- to 6-membered monocyclic ring or 8- to 10-membered bicyclic ring, which can contain 1 to 4 (1, 2, 3, or 4) atoms selected from nitrogen, oxygen, and sulfur. Preferred heteroaryls are 6- to 10-membered heteroaryls or 5- to 6-membered heteroaryls, which can contain 1, 2, or 3 atoms selected from nitrogen, oxygen, and sulfur. Examples of "heteroaryl" include, but are not limited to, furanyl, pyridyl, 2-oxo-l,2-dihydropyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzodioxolyl, benzothienyl, benzoimidazolyl, indolyl, isoindolyl, 1,3-dioxo-isoindolyl, quinolinyl, indazolyl, benzoisothiazolyl, benzoxazolyl, benzoisoxazolyl, isothiazolyl, 1H-1,2,4-triazolyl, 4H-1,2,4-triazolyl, pyridyl, pyrid-2(lH)-ononyl, pyrimidinyl, pyrazin-2(lH)-ononyl, pyrimidin-4(3H)-ononyl, pyrimidin-2(lH)-ononyl, pyridazin-3(2H)-ononyl, lH-indolyl, lH-benzo[d]imidazolyl, lH-pyrrolo[2,3-c]pyridinyl, 3H-imidazo[4,5-c]pyridinyl, isoquinolinyl, quinazolinyl, 2H-isoindolyl, furan[3,2-b]pyridinyl, furan[2,3-c]pyridinyl, thieno[2,3-c]pyridinyl, benzofuranyl, benzo[b]thiophenyl, lH-pyrrolo[3,2-b]pyridinyl, 2H-pyrrolo[3,4-c]pyridinyl,

[0139] Heteroaryl can be substituted or unsubstituted.

[0140] "Fused ring" means a polycyclic group in which two or more cyclic structures share a pair of atoms with each other, at least one of which rings has a fully conjugated pi-electron aromatic system, while 1, 2, or 3 rings can contain 0, 1, or multiple double bonds, but at least one ring does not have a fully conjugated pi-electron aromatic system, where the ring atoms are selected from 0, 1, or multiple heteroatoms selected from nitrogen, oxygen, or S(O) r (wherein r is selected from 0, 1, or 2) heteroatoms, with the remaining ring atoms being carbon. Fused rings preferably include bicyclic or tricyclic fused rings, with bicyclic fused rings preferably being the fusion of an aryl or heteroaryl group with a monocyclic heterocyclyl or monocyclic cycloalkyl group. Preferred are 6- to 14-membered, more preferably 8- to 10-membered. Examples of "fused ring" include, but are not limited to:

[0141] "Alkoxy" means a group of the formula (alkyl-O-). Alkyl is as defined above. Preferably, the alkoxy group is a C1-C6alkoxy group. Examples include, but are not limited to, methoxy, ethoxy, n-propyloxy, isopropyloxy, n-butyloxy, isobutyloxy, t-butyloxy, and the like.

[0142] "Alkylthio" means a group of the formula (alkyl-S-). Alkyl is as defined above. Preferably, the alkylthio group is a C1-C6alkylthio group. Examples include, but are not limited to, methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, isobutylthio, t-butylthio, and the like.

[0143] "Carboxy" means a group of the formula -C(=O)OH.

[0144] "Halo" means fluoro, chloro, bromo, and iodo.

[0145] "Amino" means -NH2.

[0146] "Hydroxylamino" means -NHOH.

[0147] "Cyano" means -CN.

[0148] "Benzyl" means -CH2-phenyl.

[0149] "Carboxy" means a group of the formula -C(=O)OH.

[0150] "Carboxy" means a group of the formula -C(=O)OH.

[0151] "Carboxy" means a group of the formula -C(=O)OH.

[0152] "Hydroxyalkyl" means an alkyl group as defined above which is substituted with a hydroxyl group.

[0153] "Aminoalkyl" means an alkyl group as defined above which is substituted with an amino group.

[0154] "Haloalkyl" means an alkyl group as defined above which is substituted with a halogen.

[0155] "Haloalkoxy" means an alkoxy group as defined above which is substituted with a halogen.

[0156] In the context of groups herein which refer to alkyl groups, such as alkyl, alkoxy, hydroxyalkyl, haloalkyl, haloalkoxy, alkoxy, and alkylthio, it is intended that C1-C6corresponding groups, or C1-C4corresponding groups, such as C1-C6alkyl, C1-C6haloalkyl, C1-C6haloalkoxy, C1-C6alkoxy, and C1-C6alkylthio, are preferred.

[0157] "DMSO" means dimethyl sulfoxide.

[0158] "BOC" means tert-butyloxycarbonyl.

[0159] "Bn" means benzyl.

[0160] "THP" means 2-tetrahydropyranyl.

[0161] "TFA" means trifluoroacetic acid.

[0162] "Ts" means p-toluenesulfonyl.

[0163] "Bn" means benzyl.

[0164] "SEM" means (trimethylsilyl)ethoxymethyl.

[0165] "formyl" means

[0166] "leaving group", or leaving group, is an atom or functional group that departs from a larger molecule in a chemical reaction, and is a term used in nucleophilic substitution and elimination reactions. In nucleophilic substitution reactions, the reactant that is attacked by the nucleophile is called the substrate, and the atom or group of atoms that breaks away from the substrate molecule with a pair of electrons is called the leaving group. Groups that can readily accept electrons and bear a negative charge well are good leaving groups. The smaller the pKa of the conjugate acid of the leaving group, the more readily the leaving group departs from other molecules. This is because the smaller the pKa of its conjugate acid, the greater the tendency of the corresponding leaving group to exist as an anion (or a neutral leaving group) without bonding to other atoms. Common leaving groups include, but are not limited to, halogens, mesylate, -OTs, or -OH.

[0167] "substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3, of a group are independently of one another replaced by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, which can be determined (experimentally or theoretically) by the person skilled in the art without undue effort, as possible or impossible substitutions. For example, an amino group with a free hydrogen or a hydroxyl group can be unstable when bound to a carbon atom with an unsaturated (e.g. olefinic) bond.

[0168] In the present application, "one or more" means 1 or 2 or more, for example 1, 2, 3, 4 or 5 or more, for example 1 or 2, 1 or 2 or 3, 1 or 2 or 3 or 4.

[0169] "substituted" or "substituted" as used herein, means that the group can be substituted with one or more substituents selected from the group consisting of deuterium atom, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, amino, haloalkyl, haloalkoxy, hydroxyalkyl, carboxyl, carboxylate, SF5, =0, -OR 5 , -C(=O)R 5 , -C(=O)OR 5 , -N(R 6 )C(=O)R 7 , -N(R 6 )C(=O)OR 7 , -NR 6 R 7 , -C(=O)NR 6 R 7 , -CH2NHC(=O)OR 5 , -CH2NR 6 R 7 , -S(=O) r NR 6 R 7 and -S(O) r R 5 ;

[0170] R 5 each independently is selected from the group consisting of alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further substituted with 1, 2 or 3 substituents selected from the group consisting of deuterium atom, hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -C(=O)R 8 , -C(=O)OR 8 , -OC(=O)R 8 , -NR 9 R 10 , -C(=O)NR 9 R 10 , -S(=O)2NR 9 R 10 and -N(R 9 )C(=O)R 10 ;

[0171] R 6 and R 7each independently selected from the group consisting of a hydrogen atom, a hydroxy group, an alkyl group, a cycloalkyl group, a heterocyclyl group, an aryl group, and a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclyl group, aryl group, and heteroaryl group are optionally further substituted with 1, 2, or 3 substituents selected from the group consisting of a hydroxy group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, =0, -C(=0)R 8 , -C(=0)OR 8 , -OC(=0)R 8 , -NR 9 R 10 , -C(=0)NR 9 R 10 , -S(=0)2NR 9 R 10 , and -N(R 9 )C(=0)R 10 ;

[0172] or, R 6 and R 7 , together with the atom to which they are attached, form a 4-8 membered heterocyclyl group containing 1, 2, or 3 N, O, or S(=0) r atoms, wherein said 4-8 membered heterocyclyl group is optionally further substituted with 1, 2, or 3 substituents selected from the group consisting of a hydroxy group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, =0, -C(=0)R 8 , -C(=0)OR 8 , -OC(=0)R 8 , -NR 9 R 10 , -C(=0)NR 9 R 10 , -S(=0)2NR 9 R 10 , and -N(R 9 )C(=0)R 10 ;

[0173] R 8 , R 9 , and R 10 are each independently selected from the group consisting of a hydrogen atom, an alkyl group, an amino group, a cycloalkyl group, a heterocyclyl group, an aryl group, or a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclyl group, aryl group, or heteroaryl group is optionally further substituted with 1, 2, or 3 substituents selected from the group consisting of a hydroxy group, a halogen, a nitro group, an amino group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, a carboxyl group, and a carboxylate group;

[0174] each r is independently 0, 1, or 2.

[0175] In this document, the term "wavy line" generally indicates the point of attachment of the group to the compound. The compounds of the present application can contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the present application, including but not limited to, diastereomeric, enantiomeric, and atropisomeric forms, and geometric (conformational) isomers, and mixtures thereof, such as racemates, be embraced within the scope of the present application.

[0176] Unless otherwise indicated, structures depicted throughout this specification are also meant to include all isomeric forms (e.g., diastereomeric, enantiomeric, and atropisomeric forms; for example, the R and S configurations of each asymmetric center, the (Z) and (E) double bond isomers, and the (Z) and (E) conformational isomers) of the structure; for example, the individual (R)- and (S)- configurations within each asymmetric center of the compounds described herein are within the scope of the application. Thus, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (conformational) mixtures of the present compounds are within the scope of the application.

[0177] "Pharmaceutically acceptable salt" refers to those salts which retain the biological activity of the parent compound and which are suitable for medical use. The pharmaceutically acceptable salts of the compounds of Formula (I) can be metal salts, amine salts with suitable acids.

[0178] "Pharmaceutical composition" means a mixture of one or more of the compounds described herein or physiologically acceptable salts or prodrugs thereof with other chemical components, as well as the form in which the composition is presented, for example, as a pharmaceutical formulation. The goal of a pharmaceutical composition is to facilitate administration of an active ingredient to an organism and to facilitate absorption of the active ingredient. BRIEF DESCRIPTION OF DRAWINGS

[0179] Figure 1 is a plot of tumor growth in subcutaneously xenografted tumor mice in Test Example 5 of the present application.

[0180] Figure 2 is a plot of body weight change in subcutaneously xenografted tumor mice in Test Example 5 of the present application. DETAILED DESCRIPTION

[0181] The following examples are provided to further illustrate the present application, but are not meant to limit the scope of the application.

[0182] EXAMPLE

[0183] The examples below provide the preparation of representative compounds of Formula (I) and related structural identification data. It must be understood that the following examples are illustrative only and are not meant to limit the application. 1 H NMR spectra were recorded on a Bruker instrument (400 MHz) and chemical shifts are reported in ppm using tetramethylsilane as internal standard (0.00 ppm). 1NMR notation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broad, dd = doublet of doublets, dt = doublet of triplets. If coupling constants are given, the units are Hz.

[0184] Mass spectra were determined using LC / MS instrument, and ionization mode can be ESI or APCI.

[0185] Thin layer chromatography silica gel plates were Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications of silica gel plates used in thin layer chromatography (TLC) were 0.15 mm to 0.2 mm, and the specifications of silica gel plates used in thin layer chromatography separation and purification of products were 0.4 mm to 0.5 mm.

[0186] Column chromatography generally used Yantai Huanghai silica gel 200 to 300 mesh silica gel as the carrier.

[0187] In the following examples, unless otherwise indicated, all temperatures are in degrees Celsius, and unless otherwise indicated, all starting materials and reagents were obtained from commercial suppliers or synthesized according to known methods, and all commercially available starting materials and reagents were used without further purification, unless otherwise indicated, and commercial suppliers include, but are not limited to, Aldrich Chemical Company, ABCR GmbH & Co. KG, Acros Organics, Guangzhan Chemical Technology Co., Ltd. and Jingyan Chemical Technology Co., Ltd.

[0188] CD3OD: deuterated methanol.

[0189] CDCl3: deuterated chloroform.

[0190] DMSO-d6: deuterated dimethyl sulfoxide.

[0191] DEA: diethylamine.

[0192] Ammonia: ammonia.

[0193] Argon atmosphere refers to connecting a reaction bottle to an argon balloon with a volume of about 1 L.

[0194] In the examples, unless otherwise specified, the solution in the reaction refers to an aqueous solution.

[0195] The compounds were purified by silica gel column chromatography and reverse phase column chromatography, and the eluent system was selected from: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: dichloromethane: ethyl acetate; D: trifluoroacetic acid aqueous solution and acetonitrile system. The volume ratio of the solvents is different according to the polarity of the compounds, and a small amount of acidic or basic reagent can also be added for adjustment, such as acetic acid or triethylamine, etc.

[0196] Synthesis of compound 1 in example 1

[0197] Compound 1

[0198] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0199] (S,E)-6-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-2-(1,1-Difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0200] first step

[0201] ethyl 2-acetyl-4-methylpyrimidine-5-carboxylate

[0202] 2-Acetyl-4-methylpyrimidine-5-carboxylic acid ethyl ester

[0203] To a solution of ethyl 2-chloro-4-methylpyrimidin-5-carboxylate 1a (8.97 g, 44.86 mmol) and tributyl(1-ethoxyenyl)stanane (24.30 g, 67.29 mmol) in N,N-dimethylformamide (90 mL), bis(triphenylphosphine)palladium dichloride (1.57 g, 2.24 mmol) was added. The mixture was reacted at 90 °C for 12 hours under a nitrogen atmosphere. After returning to room temperature, a saturated potassium fluoride solution (300 mL) was added to the reaction solution, followed by extraction with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was dissolved in tetrahydrofuran (150 mL). A dilute hydrochloric acid solution (2 M, 100 mL) was added, and the mixture was stirred at 25 °C for 2 hours. The reaction solution was poured into a saturated sodium bicarbonate solution (150 mL), extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated by silica gel column chromatography (eluent: system A) to give ethyl 2-acetyl-4-methylpyrimidine-5-carboxylic acid 1b (5.9 g), yield 63.2%.

[0204] MS m / z(ESI):[M+H] + =209.2

[0205] Step 2

[0206] ethyl 2-(1,1-difluoroethyl)-4-methylpyrimidine-5-carboxylate

[0207] 2-(1,1-Difluoroethyl)-4-methylpyrimidine-5-carboxylic acid ethyl ester

[0208] Ethyl 2-acetyl-4-methylpyrimidine-5-carboxylate 1b (5.9 g, 28.34 mmol) was added to diethylaminosulfur trifluoride (36.6 g, 227.08 mmol) and reacted at 25 °C for 12 hours. The reaction solution was diluted with dichloromethane (100 mL) and slowly added dropwise with saturated sodium carbonate solution (300 mL). The mixture was extracted with dichloromethane (100 mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel column chromatography (eluent: system A) to give ethyl 2-(1,1-difluoroethyl)-4-methylpyrimidine-5-carboxylate 1c (5 g), yield 76.7%.

[0209] MS m / z(ESI):[M+H] + =231.2

[0210] Step 3

[0211] ethyl 2-(1,1-difluoroethyl)-4-(2-(dimethylamino)vinyl)pyrimidine-5-carboxylate

[0212] 2-(1,1-Difluoroethyl)-4-(2-(dimethylamino)vinyl)pyrimidine-5-carboxylic acid ethyl ester

[0213] Ethyl 2-(1,1-difluoroethyl)-4-methylpyrimidine-5-carboxylic acid ester 1c (5 g, 21.72 mmol) was dissolved in N,N-dimethylformamide dimethyl acetal (40 mL) and reacted at 140 °C for 12 hours. After returning to room temperature, the reaction solution was concentrated, and the residue was separated by silica gel column chromatography (eluent: system A) to give ethyl 2-(1,1-difluoroethyl)-4-(2-(dimethylamino)yl)vinyl)pyrimidine-5-carboxylic acid ester 1d (5 g), yield 80.7%.

[0214] MS m / z(ESI):[M+H] + =286.2

[0215] Step 4

[0216] (S)-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0217] (S)-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0218] To a solution of ethyl 2-(1,1-difluoroethyl)-4-(2-(dimethylamino)vinyl)pyrimidine-5- carboxylate 1d (1 g, 3.51 mmol) in ethanol (15 mL) was added (S)-2-amino-2-cyclopropyl- ethyl-1-ol hydrochloride 1e (723.52 mg, 5.26 mmol, commercial) and acetic acid (210.49 mg, 3.51 mmol) and the reaction was stirred at 110 °C for 12 h. After cooling to room temperature, the reaction was concentrated and the residue was purified by column chromatography on silica gel (eluent: system B) to give (S)-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)- one 1f (790 mg) in 76.3% yield.

[0219] MS m / z (ESI): [M+H] + = 296.2

[0220] Fifth step

[0221] (S)-8-bromo-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0222] (S)-8-bromo-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0223] (S)-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3- d]pyrimidin-5(6H)-one

[0224] MS m / z (ESI): [M+H] + = 374.2, 376.2

[0225] Sixth step

[0226] (S)-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3- d]pyrimidin-5(6H)-one

[0227] (S)-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3- d]pyrimidin-5(6H)-one

[0228] (S)-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3- d]pyrimidin-5(6H)-one

[0229] MS m / z (ESI): [M+H] + = 372.3

[0230] Seventh step

[0231] 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-8-phenylpyrido[4,3-d]pyrimidin-6(5H)-yl)acetaldehyde

[0232] 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-8-phenylpyrido[4,3-d]pyrimidin-6(5H)-yl)acetaldehyde

[0233] To oxalyl chloride (1.34 g, 10.56 mmol) in dichloromethane (15 mL) was added a solution of dimethylsulfoxide (1.03 g, 13.19 mmol) in dichloromethane (5 mL) dropwise at -70 °C. After stirring for 30 min, a solution of (S)-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one 1h (490 mg, 1.32 mmol) in dichloromethane (5 mL) was added dropwise. After stirring for 1 h, triethylamine (2.00 g, 19.79 mmol) was added, and the reaction was allowed to warm to room temperature and stirred for 1 h. The reaction was quenched with saturated ammonium chloride solution (50 mL) and extracted with dichloromethane (25 mL x 2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography on silica gel to give 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-8-phenylpyrido[4,3-d]pyrimidin-6(5H)-yl)acetaldehyde 1i (420 mg) in 86.2% yield.

[0234] 1 H NMR (400 MHz, DMSO-d6) δ 9.73 (s, 1H), 9.69 (s, 1H), 8.33 (s, 1H), 7.71-7.68 (m, 2H), 7.52-7.48 (m, 2H), 7.44-7.41 (m, 1H), 4.44 (d, J = 10.2 Hz, 1H), 2.04 (t, J = 19.0 Hz, 3H), 1.78-1.67 (m, 1H), 0.90-0.75 (m, 2H), 0.56-0.44 (m, 2H) ppm.

[0235] Eighth Step

[0236] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0237] ​​​​​​​(E)-6-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-2-(1,1-Difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0238] Diethyl (((methanesulfonyl)methyl)phosphate 1J (299.18 mg, 1.30 mmol, prepared according to the method described in published patent WO2024010782A1)) was dissolved in tetrahydrofuran (8 mL). A solution of n-butyllithium in n-hexane (1.6 M, 595.62 μL) was slowly added dropwise at -70 °C. After stirring for 10 minutes, 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-8-phenylpyridine) was added. A solution of [4,3-d]pyrimidin-6(5H)-yl)acetaldehyde 1i (320 mg, 866.35 μmol) in tetrahydrofuran (4 mL) was reacted at -78 °C for 1 hour. Then, saturated ammonium chloride solution (30 mL) was added and stirring continued. After returning to room temperature, the mixture was extracted with ethyl acetate (20 mL × 2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative liquid chromatography (Waters 3767QDA column: XBridge C). 18 19*250mm, 10μm; mobile phase A: 10mmol / L NH4HCO3 / H2O, mobile phase B: acetonitrile; flow rate: 20mL / min), yielding (E)-6-(1-cyclopropyl-3-(methanesulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one 1k (278.7mg), yield 72.2%.

[0239] Chiral analysis methods:

[0240] Chromatographic column: 100*3mm, 3μm;

[0241] Flow rate: 1.5 mL / min;

[0242] Gradient: 10% MeOH (containing 0.1% diethylamine): 90% CO2.

[0243] The ee value of compound 1k is 7.6%.

[0244] MS m / z (ESI): 446.3 [M+1]

[0245] 1H NMR (400 MHz, MeOH-d4) δ 9.72 (s, 1H), 8.11 (s, 1H), 7.69-7.65 (m, 2H), 7.50-7.45 (m, 2H), 7.44-7.39 (m, 1H), 7.14 (dd, J = 15.3, 4.7 Hz, 1H), 6.94 (dd, J = 15.3, 1.8 Hz, 1H), 4.92 (ddd, J = 10.1, 4.6, 1.7 Hz, 1H), 3.02 (s, 3H), 2.02 (t, J = 18.6 Hz, 3H), 1.75-1.65 (m, 1H), 0.99-0.90 (m, 1H), 0.75-0.64 (m, 2H), 0.56 - 0.46 (m, 1H) ppm.

[0246] Ninth step

[0247] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0248] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0249] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one 1k (217 mg) was purified by chiral prep (column: 250*25 mm, 10 μm, flow rate 120 mL / min, gradient: 10% MeOH (containing 0.1% 7.0 mol / L of ammonia methanol solution): 90% CO2) to give the excess isomer compound (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one 1 (100.1 mg), yield 46.1%.

[0250] Chiral analysis method:

[0251] Chromatography column: 100*3 mm, 3 μm;

[0252] Flow rate: 1.5 mL / min;

[0253] Gradient: 10% MeOH (containing 0.1% diethylamine): 90% CO2.

[0254] Compound 1 retention time 2.2 min, chiral purity >99% ee.

[0255] MS m / z (ESI): 446.3 [M+1]

[0256] 1H NMR (400 MHz, MeOH-d4) δ 9.72 (s, 1H), 8.11 (s, 1H), 7.70-7.65 (m, 2H), 7.50-7.45 (m, 2H), 7.44-7.39 (m, 1H), 7.14 (dd, J = 15.3, 4.7 Hz, 1H), 6.94 (dd, J = 15.3, 1.8 Hz, 1H), 4.92 (ddd, J = 10.2, 4.7, 1.8 Hz, 1H), 3.02 (s, 3H), 2.02 (t, J = 18.6 Hz, 3H), 1.74-1.65 (m, 1H), 1.00-0.89 (m, 1H), 0.77-0.65 (m, 2H), 0.57-0.46 (m, 1H) ppm.

[0257] Compounds 2-4, 7 were synthesized according to the synthetic procedure of Reference Example 1 and their specific structures and characterization are shown in the following table:

[0258] Synthesis of compounds 1 and 1’ of Example 2

[0259] Compounds 1 and 1’

[0260] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0261] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0262] (R,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0263] (R,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8- phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0264] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8- phenylpyrido[4,3-d]pyrimidin-5(6H)-one 1k (400 mg) was purified by chiral prep (chromatographic column: 250*25 mm, 10 μm, flow rate 50 mL / min, gradient: 25% isopropanol: 75% CO2), to give (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8- phenylpyrido[4,3-d]pyrimidin-5(6H)-one 1 (173 mg), 43.3% yield and (R,E)-6-(1- cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenylpyrido[4,3- d]pyrimidin-5(6H)-one 1' (140 mg), 35.0% yield.

[0265] Chiral analysis method:

[0266] Chromatographic column: 100*3 mm, 3 μm;

[0267] Flow rate: 1.5 mL / min;

[0268] Gradient: 10% MeOH (with 0.1% diethylamine): 90% CO2.

[0269] Compound 1 retention time 2.2 min, chiral purity >99% ee.

[0270] MS m / z (ESI): 446.3 [M+1]

[0271] 1H NMR (400MHz, DMSO-d6) δ9.68 (s, 1H), 8.34 (s, 1H), 7.69 (d, J = 7.4Hz, 2H), 7.50 (t, J=7.4Hz,2H),7.42(t,J=7.3Hz,1H),7.06(dd,J=15.3,4.8Hz,1H),6.96(d,J=15.5 Hz,1H),4.87–4.76(m,1H),3.03(s,3H),2.04(t,J=19.0Hz,3H),1.89–1.79(m,1H ),0.87–0.77(m,1H),0.74–0.66(m,1H),0.63–0.54(m,1H),0.45–0.35(m,1H)ppm.

[0272] Compound 1' has a retention time of 3.0 minutes and a chiral purity of 97% ee.

[0273] MS m / z (ESI): 446.3 [M+1]

[0274] 1 H NMR(400MHz,MeOH-d4)δ9.72(s,1H),8.11(s,1H),7.70–7.65(m,2H),7.51– 7.45(m,2H),7.44–7.39(m,1H),7.14(dd,J=15.3,4.6Hz,1H),6.94(dd,J=15 .3,1.7Hz,1H),4.94–4.92(m,1H),3.02(s,3H),2.02(t,J=18.6Hz,3H),1.7 4–1.65(m,1H),0.99–0.91(m,1H),0.75–0.65(m,2H),0.56–0.47(m,1H)ppm.

[0275] Example 3 Synthesis of compounds 2k, 2 and 2'

[0276] Compounds 2k, 2 and 2'

[0277] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0278] (E)-2-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-6-(1,1-Difluoroethyl)-4-phenyl-2,7-naphthidium-1(2H)-one

[0279] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0280] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0281] (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0282] (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0283] First step

[0284] methyl 6-chloro-4-methylnicotinate

[0285] methyl 6-chloro-4-methylnicotinate

[0286] Methyl 6-chloro-4-methylnicotinate 2b (32 g, 65.7% yield) was prepared according to the following procedure. 6-Chloro-4-methyl nicotinic acid 2a (45 g, 262.27 mmol, commercially available) was dissolved in phosphorus oxychloride (300 mL) and reacted at 100 °C for 16 hours. The reaction was cooled to room temperature and concentrated under reduced pressure. The residue was slowly added to methanol (21 mL) at 0 °C and stirred for 2 hours at 0 °C. The solvent was removed under reduced pressure and the residue was dissolved in dichloromethane (200 mL). The solution was added to saturated sodium bicarbonate solution (500 mL) and the organic phase was separated. The aqueous phase was extracted with dichloromethane (200 mL x 2) and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by column chromatography on silica gel (eluent: A system) to give methyl 6-chloro-4-methylnicotinate 2b (32 g, 65.7% yield).

[0287] MS m / z (ESI): [M+H] + = 186.2

[0288] Second step

[0289] methyl 4-methyl-6-(prop-1-en-2-yl)nicotinate

[0290] 4-methyl-6-(prop-1-en-2-yl)nicotinic acid methyl ester

[0291] Methyl 6-acetyl-4-methylnicotinate

[0292] MS m / z (ESI): [M+H] + = 192.0

[0293] Third step

[0294] methyl 6-acetyl-4-methylnicotinate

[0295] 6-acetyl-4-methyl-nicotinic acid methyl ester

[0296] Methyl 6-acetyl-4-methylnicotinate

[0297] MS m / z (ESI): [M+H] + = 194.0

[0298] Fourth step

[0299] methyl 6-(1,1-difluoroethyl)-4-methylnicotinate

[0300] 6-(1,1-difluoroethyl)-4-methylnicotinic acid methyl ester

[0301] Methyl 6-acetyl-4-methylnicotinate 2d (22 g, 113.87 mmol) was dissolved in diethylamine sulfide (80 mL) and reacted at 50 °C for 16 h. After returning to room temperature, the reaction solution was diluted with dichloromethane (300 mL), added to saturated sodium bicarbonate solution (600 mL), extracted with dichloromethane (300 mL x 2), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain methyl 6-(1,1-difluoroethyl)-4-methylnicotinate 2e (20 g) with a yield of 81.6%.

[0302] MS m / z (ESI): [M+H] + = 216.1

[0303] Fifth step

[0304] 6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one

[0305] 6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one

[0306] Under a nitrogen environment, 4A molecular sieves (2 g) were added to a reaction bottle, activated at high temperature and cooled to room temperature, then methyl 6-(1,1-difluoroethyl)-4-methylnicotinate 2e (5 g, 23.23 mmol) and dimethyl sulfoxide (80 mL) were added, followed by the addition of 1,3,5-triazine (2.26 g, 27.88 mmol), potassium tert-butoxide in tetrahydrofuran (1 M, 30.20 mL), and reacted at 80 °C for 2 h. After returning to room temperature, water (50 mL) was added, extracted with ethyl acetate (50 mL x 3), the combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated, and the obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain 6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one 2f (2.2 g) with a yield of 45.1%.

[0307] MS m / z (ESI): [M+H] + = 211.1

[0308] Sixth step

[0309] ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-2,7-naphthyridin-2(1H)-yl)acetate

[0310] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-2,7-naphthyridin-2(1H)-yl)acetic acid ethyl ester

[0311] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-2,7-naphthyridin-2(1H)-yl)acetic acid ethyl ester

[0312] MS m / z (ESI): [M+H] + = 337.2

[0313] Seventh step

[0314] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one

[0315] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one

[0316] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one

[0317] MS m / z (ESI): [M+H] + = 295.0

[0318] Eighth Step

[0319] 4-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one

[0320] 4-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one

[0321] 4-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-2,7-naphthyridin-1(2H)-one

[0322] MS m / z (ESI): [M+H] + = 373.0 / 375.0

[0323] Ninth Step

[0324] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0325] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0326] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-oneUnder nitrogen atmosphere, 4-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1- difluoroethyl)-2,7-naphthyridin-1(2H)-one 2i (1.72 g, 4.61 mmol) was dissolved in a mixed solvent of 1,4-dioxane (20 mL) and water (5 mL), and benzeneboronic acid (842.94 mg, 6.91 mmol), potassium carbonate (1.91 g, 13.83 mmol), [1,1'- bis(diphenylphosphino)ferrocene]dichloropalladium (341.88 mg, 460.89 µmol) were added, and the reaction was carried out at 100°C for 2 hours. After being returned to room temperature, the reaction solution was filtered with diatomite, and the filter cake was washed with ethyl acetate. The combined organic phase was concentrated under reduced pressure, and the obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one 2j (1.48 g) with a yield of 86.7%.

[0327] MS m / z (ESI): [M+H] + = 371.1

[0328] Tenth step

[0329] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenyl-2,7-naphthyridin-2(1H)-yl)acetaldehyde

[0330] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenyl-2,7-naphthyridin-2(1H)-yl)acetaldehyde

[0331] Oxalyl chloride (2.74 g, 21.60 mmol) was added to dichloromethane (20 mL) and cooled to -78°C, and slowly added dropwise to a dimethyl sulfoxide (2.11 g, 27.00 mmol) solution in dichloromethane (10 mL) while stirring at -78°C for 10 minutes. Then, 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one 2j (1 g, 2.70 mmol) in dichloromethane (5 mL) was continuously added dropwise, and the reaction was continued for 1 hour. Then, triethylamine (4.10 g, 40.50 mmol, 5.62 mL) was added dropwise, and the reaction was continued at room temperature for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (10 mL), and extracted with dichloromethane (10 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenyl-2,7-naphthyridin-2(1H)-yl)acetaldehyde 2m (1 g), which was directly used in the next step.

[0332] MS m / z (ESI): [M+H] + = 369.1

[0333] Tenth step

[0334] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0335] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0336] Under nitrogen atmosphere, diethyl ((methylsulfonyl)methyl)phosphonate 1j (937.43 mg, 4.07 mmol) was dissolved in tetrahydrofuran (10 mL) and cooled to -78 °C. n-Butyllithium solution in n-hexane (1.6 M, 2.21 mL) was added and the reaction was continued at -78 °C for 1 hour. A solution of 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenyl-2,7-naphthyridin-2(1H)-yl)acetaldehyde 2m (1 g, 2.71 mmol) in tetrahydrofuran (10 mL) was added and the reaction was continued at -78 °C for 1 hour. The reaction was quenched by adding saturated ammonium chloride solution (10 mL) and the temperature was allowed to return to room temperature. The organic phase was extracted with ethyl acetate (10 mL x 3), the combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue obtained was purified by preparative liquid chromatography (Gilson GX-281 column: SunFire C 18 , 19*250 mm, 10 μm; mobile phase A: 0.1% formic acid in water, mobile phase B: acetonitrile; flow rate: 25 mL / min) to give (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one 2k (309.90 mg) in 25.7% yield.

[0337] MS m / z (ESI): [M+H] + = 445.1

[0338] 1H NMR (400 MHz, DMSO-d6) δ 9.51 (s, 1H), 8.00 (s, 1H), 7.63 (s, 1H), 7.60 - 7.54 (m, 2H), 7.54 - 7.47 (m, 3H), 7.04 (dd, J = 15.3, 4.9 Hz, 1H), 6.93 (dd, J = 15.4, 1.2 Hz, 1H), 4.83 (dd, J = 9.6, 4.6 Hz, 1H), 3.03 (s, 3H), 2.01 (t, J = 19.2 Hz, 3H), 1.85 - 1.71 (m, 1H), 0.88 - 0.71 (m, 1H), 0.71 - 0.62 (m, 1H), 0.61 - 0.50 (m, 1H), 0.45 - 0.32 (m, 1H) ppm.

[0339] Twelfth step

[0340] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0341] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one

[0342] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one 2k (300 mg) was purified by chiral prep (Chiralpak® IC column: 250*30 mm 10 pm, flow rate 120 mL / min, gradient: 35% MeOH:65% CO2), to give (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one 2 (129 mg), 43% yield, and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenyl-2,7-naphthyridin-1(2H)-one 2' (132 mg), 44% yield. Chiral analytical method:

[0343] Chromatography column: 100*3 mm, 3 pm;

[0344] Flow rate: 2.0 mL / min;

[0345] Gradient: 80% MeOH (with 0.1% diethylamine):20% CO2.

[0346] Compound 2 retention time 2.84 min, chiral purity >99% ee.

[0347] MS m / z (ESI): [M+H]+ = 445.1 + = 445.1

[0348] 1 H NMR (400 MHz, DMSO-d6) δ 9.51 (s, 1H), 7.99 (s, 1H), 7.63 (s, 1H), 7.60 - 7.55 (m, 2H), 7.53 - 7.47 (m, 3H), 7.04 (dd, J = 15.4, 4.8 Hz, 1H), 6.93 (d, J = 15.4 Hz, 1H), 4.83 (dd, J = 9.5, 5.2 Hz, 1H), 3.03 (s, 3H), 2.01 (t, J = 19.2 Hz, 3H), 1.83 - 1.72 (m, 1H), 0.84 - 0.74 (m, 1H), 0.71 - 0.63 (m, 1H), 0.62 - 0.52 (m, 1H), 0.43 - 0.31 (m, 1H) ppm.

[0349] Compound 2' retention time 1.69 min, chiral purity 98.6% ee.

[0350] MS m / z (ESI): [M+H]+ = 445.1

[0351] 1H NMR (400 MHz, DMSO-d6) δ 9.51 (s, 1H), 8.00 (s, 1H), 7.63 (s, 1H), 7.60 - 7.55 (m, 2H), 7.54 - 7.47 (m, 3H), 7.04 (dd, J = 15.3, 4.9 Hz, 1H), 6.93 (dd, J = 15.3, 1.3 Hz, 1H), 4.83 (dd, J = 9.2, 4.7 Hz, 1H), 3.03 (s, 3H), 2.01 (t, J = 19.2 Hz, 3H), 1.84 - 1.69 (m, 1H), 0.83 - 0.74 (m, 1H), 0.71 - 0.62 (m, 1H), 0.60 - 0.51 (m, 1H), 0.42 - 0.30 (m, 1H) ppm.

[0352] Synthesis of compounds 3k, 3 and 3'

[0353] Compound 3k, 3 and 3'

[0354] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0355] (E)-6-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-2-(1,1-Difluoroethyl)-8-phenyl-1,6-naphthidium-5(6H)-one

[0356] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0357] (S,E)-6-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-2-(1,1-Difluoroethyl)-8-phenyl-1,6-naphthidium-5(6H)-one

[0358] (R,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0359] (R,E)-6-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-2-(1,1-Difluoroethyl)-8-phenyl-1,6-naphthidin 5(6H)-one

[0360] first step

[0361] methyl 6-acetyl-2-methylnicotinate

[0362] 6-Acetyl-2-methylnicotinic acid methyl ester

[0363] Methyl 6-(1,1-difluoroethyl)-2-methylnicotinate + = 194.2

[0364] Second step

[0365] Methyl 6-(1,1-difluoroethyl)-2-methylnicotinate

[0366] Methyl 6-(1,1-difluoroethyl)-2-methylnicotinate

[0367] Methyl 6-(1,1-difluoroethyl)-2-methylnicotinate + = 216.2

[0368] Third step

[0369] 2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0370] 2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0371] Methyl 6-(1,1-difluoroethyl)-2-methylnicotinate 3c (1.3 g, 6.04 mmol) was added to dimethyl sulfoxide (15 mL), 1,3,5-triazine (587.77 mg, 7.25 mmol) and potassium tert-butoxide in tetrahydrofuran (1 M, 7.85 mL) were added, and the reaction was heated to 80 °C for 12 h. The reaction was cooled to room temperature, water (40 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by column chromatography on silica gel (eluent: system A) to give 2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 3d (1.1 g) in 86.6% yield.

[0372] MS m / z (ESI): [M+H] + = 211.1

[0373] Fourth step

[0374] ethyl 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-1,6-naphthyridin-6(5H)-yl)acetate

[0375] 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-1,6-naphthyridin-6(5H)-yl)acetic acid

[0376] Methyl 6-(1,1-difluoroethyl)-2-methylnicotinate 3c (1.3 g, 6.04 mmol) was added to dimethyl sulfoxide (15 mL), 1,3,5-triazine (587.77 mg, 7.25 mmol) and potassium tert-butoxide in tetrahydrofuran (1 M, 7.85 mL) were added, and the reaction was heated to 80 °C for 12 h. The reaction was cooled to room temperature, water (40 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by column chromatography on silica gel (eluent: system A) to give 2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 3d (1.1 g) in 86.6% yield.

[0377] MS m / z (ESI): [M+H] + = 337.3

[0378] Fifth step

[0379] 6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-1,6-naphthyridin-5(6H)-one

[0380] 6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-1,6-naphthyridin-5(6H)-one

[0381] Ethyl 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-1,6-naphthyridin-6(5H)-yl)acetate 3e (1.1 g, 3.27 mmol) was added to tetrahydrofuran (10 mL), cooled to 0 °C, lithium borohydride tetrahydrofuran solution (2 M, 1.64 mL) was added, and the reaction was allowed to proceed at room temperature for 12 hours. Saturated ammonium chloride solution (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by column chromatography on silica gel (eluent: A system) to give 6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-1,6-naphthyridin-5(6H)-one 3f (500 mg) in a yield of 52.0%.

[0382] MS m / z (ESI): [M+H] + = 295.2

[0383] Sixth step

[0384] 8-bromo-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-1,6-naphthyridin-5(6H)-one

[0385] 8-bromo-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-1,6-naphthyridin-5(6H)-one

[0386] Ethyl 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-1,6-naphthyridin-6(5H)-yl)acetate 3e (1.1 g, 3.27 mmol) was added to tetrahydrofuran (10 mL), cooled to 0 °C, lithium borohydride tetrahydrofuran solution (2 M, 1.64 mL) was added, and the reaction was allowed to proceed at room temperature for 12 hours. Saturated ammonium chloride solution (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by column chromatography on silica gel (eluent: A system) to give 6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-1,6-naphthyridin-5(6H)-one 3f (500 mg) in a yield of 52.0%.

[0387] MS m / z (ESI): [M+H] + = 374.9

[0388] Seventh step

[0389] 6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0390] 6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0391] 8-bromo-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-1,6-naphthyridin-5(6H)-one 3g (600 mg, 1.61 mmol), phenylboronic acid (294.05 mg, 2.41 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL), potassium carbonate (666.60 mg, 4.82 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (117.5 mg, 160.78 μmol) were added, and the reaction was carried out at 100°C for 2 hours. After being restored to room temperature, the filter cake was filtered with diatomite, washed with ethyl acetate (10 mL), the filtrate was collected, water (10 mL) was added, and ethyl acetate (10 mL x 3) was extracted. The combined organic layer was dried with anhydrous sodium sulfate, filtered and concentrated. The obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain 6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one 3h (492.2 mg), with a yield of 82.7%.

[0392] MS m / z (ESI): [M+H] + = 371.2

[0393] Eighth step

[0394] 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-8-phenyl-1,6-naphthyridin-6(5H)-yl)acetaldehyde

[0395] 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-8-phenyl-1,6-naphthyridin-6(5H)-yl)acetaldehyde

[0396] Oxalyl chloride (1.01 g, 7.99 mmol) was added into dichloromethane (2 mL) and cooled to -78 °C, dimethyl sulfoxide (780.51 mg, 9.99 mmol) was added dropwise, after keeping -78 °C for 10 min, 6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one 3h (370 mg, 998.94 μmol) in dichloromethane (1 mL) was added dropwise, after keeping -78 °C for 1 h, triethylamine (1.82 g, 17.98 mmol) was added slowly dropwise, the reaction was continued to stir at room temperature for 12 h. Saturated ammonium chloride solution (10 mL) was used to quench the reaction, ethyl acetate (10 mL x 3) was used to extract the product, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-8-phenyl-1,6-naphthyridin-6(5H)-yl)acetaldehyde 3i (400 mg), which was used directly in the next step.

[0397] Ninth step

[0398] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0399] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0400] ((Methy lsulfony l)methyl)phosphonic acid diethyl ester 1j (346.85 mg, 1.51 mmol) was dissolved in tetrahydrofuran (6.70 mL) under nitrogen atmosphere, cooled to -78 °C, n-butyllithium in n-hexane (1.6 M, 816.08 μί) was added, after 1 hour of reaction at -78 °C, 2-cyclopropyl-2-(2-(1,1-difluoroethyl)-5-oxo-8-phenyl-1,6-naphthyridin-6(5H)-yl)acetaldehyde 3i (370 mg, 1.00 mmol) in tetrahydrofuran (6.70 mL) was added, after 1 hour of reaction at -78 °C, saturated ammonium chloride solution (10 mL) was added to quench, it was allowed to warm to room temperature, extracted with ethyl acetate (30 mL x 3), the combined organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, the obtained residue was purified by preparative liquid chromatography (Gilson GX-281 column: Agilent pursuit XRS C18, 21.2*250 mm, 10 μιη; mobile phase A: 0.1% formic acid in water, mobile phase B: acetonitrile; flow rate: 25 mL / min), (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one 3k (116 mg) was obtained in 26.1% yield.

[0401] MS m / z (ESI): [M+H] + = 445.1

[0402] Tenth step

[0403] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0404] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one

[0405] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6-naphthyridin-5(6H)-one 3k (116 mg) was purified by chiral preparation (Chiral column: 250*30 mm 10 pm, flow rate 120 mL / min, gradient: 30% MeOH: 70% CO2) to give (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8- phenyl-1,6-naphthyridin-5(6H)-one 3 (31.6 mg), 27.2% yield, and (R,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)-8-phenyl-1,6- naphthyridin-5(6H)-one 3' (35.0 mg), 30.2% yield. Chiral analysis method:

[0406] Column: 100*3 mm, 3 pm;

[0407] Flow rate: 1.5 mL / min;

[0408] Gradient: 25% MeOH: 75% CO2.

[0409] Compound 3 retention time 1.96 min, chiral purity >99% ee.

[0410] MS m / z (ESI): [M+H] + = 445.2

[0411] 1 H NMR (400 MHz, MeOH-d4) δ 8.90 (d, J = 8.3 Hz, 1H), 7.92 (d, J = 8.4 Hz, 1H), 7.87 (s, 1H), 7.70 (d, J = 7.3 Hz, 2H), 7.50 (t, J = 7.5 Hz, 2H), 7.44 (dd, J = 8.6, 5.9 Hz, 1H), 7.19 (dd, J = 15.3, 4.3 Hz, 1H), 6.94 (dd, J = 15.3, 1.6 Hz, 1H), 5.01 - 4.96 (m, 1H), 3.06 (s, 3H), 2.01 (t, J = 18.9 Hz, 3H), 1.79 - 1.67 (m, 1H), 1.05 - 0.92 (m, 1H), 0.78 - 0.66 (m, 2H), 0.61 - 0.48 (m, 1H) ppm. Compound 3' retention time 1.13 min, chiral purity >99% ee.

[0412] MS m / z (ESI): [M+H] + = 445.1

[0413] 1H NMR (400 MHz, DMSO) δ 8.82 (d, J = 8.3 Hz, 1H), 8.05 (s, 1H), 7.89 (d, J = 8.3 Hz, 1H), 7.73 - 7.66 (m, 2H), 7.48 (t, J = 7.5 Hz, 2H), 7.40 (t, J = 7.3 Hz, 1H), 7.06 (dd, J = 15.4, 4.9 Hz, 1H), 6.93 (dd, J = 15.3, 1.4 Hz, 1H), 4.89 - 4.79 (m, 1H), 3.03 (s, 3H), 2.00 (t, J = 19.2 Hz, 3H), 1.87 - 1.76 (m, 1H), 0.85 - 0.75 (m, 1H), 0.73 - 0.63 (m, 1H), 0.63 - 0.52 (m, 1H), 0.42 - 0.33 (m, 1H) ppm.

[0414] Synthesis of compounds 5k, 5 and 5' of Example 5

[0415] Compound 5k

[0416] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0417] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0418] Compound 5 and 5'

[0419] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one and (R,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0420] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8- phenylpyrido [4,3-d] pyrimidin-5(6H)-one and (R,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8- phenylpyrido [4,3-d] pyrimidin-5(6H)-one

[0421] First step

[0422] ethyl 2-cyano-2-cyclopropylacetate

[0423] 2-cyclopropylacetonitrile

[0424] Diethyl carbonate (14.56 g, 123.28 mmol) was dissolved in toluene (50 mL), sodium hydride (5.42 g, 135.61 mmol, 60% purity) was added portionwise, the temperature was raised to 110 °C, a solution of 2-cyclopropylacetonitrile 5a (5 g, 61.64 mmol, commercially available) in toluene (15 mL) was added dropwise slowly, and the reaction was carried out at 120 °C for 18 hours. It was brought to room temperature, glacial acetic acid (30 mL) was added, ice water (100 mL) was added, ethyl acetate (70 mL x 3) was extracted, the organic phases were combined, concentrated under reduced pressure, dissolved in acetonitrile (50 mL), n-hexane (100 mL) was added, the phases were separated, the acetonitrile phase was collected, and concentrated under reduced pressure to obtain ethyl 2-cyano-2-cyclopropylacetate 5b (4.55 g) with a yield of 48.2%.

[0425] 1 H NMR (400 MHz, DMSO) δ 4.20 (q, J = 7.1 Hz, 2H), 3.88 (d, J = 8.4 Hz, 1H), 1.35 - 1.28 (m, 1H), 1.23 (t, J = 7.1 Hz, 3H), 0.69 - 0.59 (m, 2H), 0.47 - 0.37 (m, 2H).

[0426] Second step

[0427] methyl 2-(cyclopropanecarbonyl)-4-methylpyrimidine-5-carboxylate

[0428] 2-(cyclopropanecarbonyl)-4-methylpyrimidine-5-carboxylic acid methyl ester

[0429] Ethyl 2-cyano-2-cyclopropylacetate 5b (500 mg, 3.26 mmol) and ethyl 2-chloro-4- methylpyrimidine-5-carboxylate (654.87 mg, 3.26 mmol) were dissolved in N,N- dimethylformamide (20 mL) under nitrogen protection, cesium carbonate (3.19 g, 9.79 mmol) and 4A molecular sieves (100 mg) were added, after reaction at room temperature for 12 hours, oxygen was replaced to continue to react for 3 hours. The reaction solution was diluted with water (80 mL), filtered, the filtrate was extracted with dichloromethane (40 mL x 3), the combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated, the obtained residue was purified by silica gel column chromatography (eluent: A system) to obtain methyl 2-(cyclopropylcarbonyl)-4-methylpyrimidine-5-carboxylate 5c (139 mg) with a yield of 18.2%.

[0430] MS m / z (ESI): [M+H] + = 271.1

[0431] The compound of racemic 5k (300 mg) was synthesized using 5c according to the method of Example 1.

[0432] SFC resolution

[0433] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one and (R,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0434] (S,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one and (R,E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8-phenylpyrido[4,3-d]pyrimidin-5(6H)-one

[0435] (E)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(cyclopropyldifluoromethyl)-8- phenylpyrido[4,3-d]pyrimidin-5(6H)-one 5k (300 mg) was purified by chiral preparation (chiral column: 250*30 mm 10 μm, flow rate 120 mL / min, gradient: 37% MeOH: 63% CO2) to give the longer-retained single configuration compound 5 (115 mg, 38.3% yield) and the shorter-retained single configuration compound 5' (114 mg, 38% yield).

[0436] Chiral analysis method

[0437] Chromatography column: 100*3 mm, 3 μm;

[0438] Flow rate: 2 mL / min;

[0439] Gradient: 65% MeOH (with 0.1% diethylamine): 35% CO2.

[0440] Compound 5 retention time 1.7 min, chiral purity >99% ee.

[0441] MS m / z (ESI): [M+H] + = 472.2

[0442] 1 H NMR (400 MHz, DMSO) δ 9.68 (s, 1H), 8.33 (s, 1H), 7.72 - 7.68 (m, 2H), 7.52 - 7.47 (m, 2H), 7.43 - 7.39 (m, 1H), 7.06 (dd, J = 15.3, 4.6 Hz, 1H), 6.96 (dd, J = 15.3, 1.3 Hz, 1H), 4.82 (dd, J = 9.3, 4.6 Hz, 1H), 3.03 (s, 3H), 1.90 - 1.79 (m, 2H), 0.84 - 0.77 (m, 1H), 0.69 - 0.72 (m, 5H), 0.63 - 0.55 (m, 1H), 0.43 - 0.36 (m, 1H) ppm.

[0443] Compound 5' retention time 1.1 min, chiral purity 98.3% ee.

[0444] MS m / z (ESI): [M+H] + = 472.2

[0445] 1H NMR (400 MHz, DMSO) δ 9.68 (s, 1H), 8.33 (s, 1H), 7.72 - 7.68 (m, 2H), 7.51 - 7.47 (m, 2H), 7.43 - 7.39 (m, 1H), 7.06 (dd, J = 15.3, 4.6 Hz, 1H), 6.96 (dd, J = 15.4, 1.3 Hz, 1H), 4.82 (dd, J = 10.0, 4.3 Hz, 1H), 3.02 (s, 3H), 1.89 - 1.79 (m, 2H), 0.84 - 0.78 (m, 1H), 0.70 (d, J = 6.5 Hz, 5H), 0.62 - 0.55 (m, 1H), 0.43 - 0.37 (m, 1H) ppm.

[0446] Synthesis of compounds 10k, 10 and 10' of Example 6

[0447] Compound 10k

[0448] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one

[0449] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one

[0450] Compound 10 and 10'

[0451] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one

[0452] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one

[0453] First step

[0454] 6-(1-ethoxyvinyl)isoquinolin-1(2H)-one

[0455] 6-(1-ethoxyvinyl)isoquinolin-1(2H)-one

[0456] 6-(1-ethoxyvinyl)isoquinolin-1(2H)-one

[0457] Second step

[0458] 6-acetylisoquinolin-1(2H)-one

[0459] 6-acetylisoquinolin-1(2H)-one

[0460] 6-acetylisoquinolin-1(2H)-one

[0461] MS m / z (ESI): [M+H] + = 188.2

[0462] Third step

[0463] ethyl 2-(6-acetyl-1-oxoisoquinolin-2(1H)-yl)-2-cyclopropylacetate

[0464] 2-(6-acetyl-1-oxoisoquinolin-2(1H)-yl)-2-cyclopropylacetate

[0465] Ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxoisoquinolin-2(1H)- yl)acetate + = 314.2

[0466] Fourth step

[0467] ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxoisoquinolin-2(1H)- yl)acetate

[0468] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxoisoquinolin-2(1H)-yl)acetic acid

[0469] Ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxoisoquinolin-2(1H)- yl)acetate

[0470] MS m / z (ESI): [M+H] + = 336.3

[0471] Fifth step

[0472] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)isoquinolin-1(2H)-one

[0473] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)isoquinolin-1(2H)-one

[0474] Ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxoisoquinolin-2(1H)-yl)acetate 10e (0.5 g, 1.49 mmol) was dissolved in tetrahydrofuran (6 mL) under nitrogen protection, cooled to 0 °C, and a tetrahydroaluminum lithium tetrahydrofuran solution (2.5 M, 298.20 μL) was added. The reaction was carried out at room temperature for 2 hours. The reaction solution was quenched with 10% sodium hydroxide solution (3 mL), extracted with ethyl acetate (10 mL x 3), and the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The resulting residue was separated by silica gel column chromatography (eluent: A system) to obtain 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)isoquinolin-1(2H)-one 10f (206 mg) with a yield of 47.1%.

[0475] MS m / z (ESI): [M+H] + = 294.2

[0476] Step 6

[0477] 4-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)isoquinolin-1(2H)-one

[0478] 4-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)isoquinolin-1(2H)-one

[0479] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)isoquinolin-1(2H)-one 10f (365 mg, 1.24 mmol) was dissolved in acetonitrile (5 mL), and N-bromosuccinimide (232.56 mg, 1.31 mmol) was added. The reaction was carried out at 80 °C for 12 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was separated by silica gel column chromatography (eluent: A system) to obtain 4-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)isoquinolin-1(2H)-one 10g (274 mg) with a yield of 59.2%.

[0480] MS m / z (ESI): [M+H] + = 372.1 / 374.1

[0481] Step 7

[0482] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one

[0483] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one

[0484] After 4-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl) isoquinolin-1(2H)-one 10g (254mg, 682.42μmol) was dissolved in 1,4-dioxane (5mL) and water (1mL), potassium carbonate (235.79mg, 1.71mmol) and 1,1'-bis(diisopropylphosphino) ferrocene palladium dichloride (50.62mg, 68.24μmol) were added, and the reaction was carried out at 100°C for 2 hours. After being restored to room temperature, the reaction solution was added with water (20mL), and the mixture was extracted with ethyl acetate (10mL x 3). The combined organic phase was washed with saturated sodium chloride solution (10mL), dried over anhydrous sodium sulfate, concentrated by filtration, and the obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one 10h (219mg), with a yield of 86.9%.

[0485] MS m / z (ESI): [M+H] + = 370.3

[0486] Eighth step

[0487] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylisoquinolin-2(1H)-yl)acetaldehyde

[0488] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylisoquinolin-2(1H)-yl)acetaldehyde

[0489] Oxalyl chloride (601.98 mg, 4.74 mmol) was dissolved in dichloromethane (2 mL) and cooled to -78 °C. Dimethyl sulfoxide (463.21 mg, 5.93 mmol) was added dropwise to the solution and stirred at -78 °C for 30 min. A solution of 2-(l-cyclopropyl-2-hydroxyethyl)-6-(l,l-difluoroethyl)-4- phenylisoquinolin-l(2H)-one 10h (219 mg, 592.85 μmol) in dichloromethane (2 mL) was added dropwise to the solution and the reaction was continued for 1 h. Triethylamine (899.84 mg, 8.89 mmol) was added dropwise and the reaction was continued for 1 h at room temperature. The reaction was diluted with water (20 mL) and extracted with dichloromethane (20 mL x 3). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 2-cyclopropyl-2-(6-(l,l-difluoroethyl)-l-oxo-4-phenylisoquinolin-2(lH)-yl)acetaldehyde 10i (210 mg), which was used directly in the next step.

[0490] MS m / z (ESI): [M+H] + = 368.3

[0491] Ninth step

[0492] (E)-2-(l-cyclopropyl-3-(methylsulfonyl)allyl)-6-(l,l-difluoroethyl)-4-phenylisoquinolin-l(2H)-one

[0493] (E)-2-(l-cyclopropyl-3-(methylsulfonyl)allyl)-6-(l,l-difluoroethyl)-4-phenylisoquinolin-l(2H)-one

[0494] ((Methylsulfonyl)methyl)phosphonic acid diethyl ester lj (108.09 mg, 469.53 μmol) was dissolved in tetrahydrofuran (2 mL) and cooled to -78 °C. n-Butyllithium solution in n-hexane (1.6 mol / L, 254.33 μL) was added dropwise to the solution and the reaction was continued for 30 min at -78 °C. A solution of 2-cyclopropyl-2-(6-(l,l-difluoroethyl)-l-oxo-4-phenylisoquinolin-2(lH)-yl)acetaldehyde 10i (115 mg, 313.02 μmol) in tetrahydrofuran (2 mL) was added dropwise to the solution and the reaction was continued for 20 min at -78 °C. The reaction was quenched by adding saturated ammonium chloride solution (5 mL) and diluted with water (10 mL). The reaction was extracted with dichloromethane (10 mL x 3). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography (Waters 3767 / Qda, column: XBridge C 18,19*250mm,10μm; mobile phase A: 10mmol / L ammonium bicarbonate aqueous solution, mobile phase B: acetonitrile; flow rate: 20 mL / min) to give (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one 10k (44.63 mg) in 32.2% yield.

[0495] MS m / z (ESI): [M+H] + = 444.0

[0496] 1 H NMR (400 MHz, DMSO) δ 8.45 (d, J = 8.5 Hz, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.67 (d, J = 6.5 Hz, 2H), 7.58 - 7.53 (m, 2H), 7.52 - 7.45 (m, 3H), 7.03 (dd, J = 15.3, 5.0 Hz, 1H), 6.89 (dd, J = 15.3, 1.5 Hz, 1H), 4.85 (dd, J = 9.3, 5.7 Hz, 1H), 3.03 (s, 3H), 1.95 (t, J = 19.0 Hz, 3H), 1.80 - 1.71 (m, 1H), 0.81 - 0.73 (m, 1H), 0.68 - 0.61 (m, 1H), 0.60 - 0.51 (m, 1H), 0.39 - 0.31 (m, 1H) ppm.

[0497] SFC resolution

[0498] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one

[0499] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylisoquinolin-1(2H)-one

[0500] (E)-2-(l-cyclopropyl-3-(methylsulfonyl)allyl)-6-(l,l-difluoroethyl)-4- phenylisoquinolin-l(2H)-one 10k (650 mg) was purified by chiral prep (Chiralpak® OJ column: 250*30 mm 10 μm, flow rate 140 mL / min, gradient: 50% MeOH: 50% CO2) to give the longer ret time single enantiomer compound 10 (227 mg, 34.9% yield) and the shorter ret time single enantiomer compound 10' (242 mg, 37.2% yield).

[0501] Chiral analysis method

[0502] Chromatography column: 100*4.6 mm, 5 μm;

[0503] Flow rate: 3 mL / min;

[0504] Gradient: 30% MeOH: 70% CO2.

[0505] Compound 10 ret time 3.0 min, chiral purity >99% ee.

[0506] MS m / z (ESI): [M+H] + = 444.0

[0507] 1 H NMR (400 MHz, DMSO-d6) δ 8.45 (d, J = 8.4 Hz, 1H), 7.77 - 7.72 (m, 1H), 7.67 (d, J = 6.4 Hz, 2H), 7.59 - 7.53 (m, 2H), 7.53 - 7.45 (m, 3H), 7.03 (dd, J = 15.3, 5.1 Hz, 1H), 6.89 (dd, J = 15.3, 1.5 Hz, 1H), 4.88 - 4.83 (m, 1H), 3.03 (s, 3H), 1.95 (t, J = 19.0 Hz, 3H), 1.81 - 1.70 (m, 1H), 0.81 - 0.73 (m, 1H), 0.68 - 0.61 (m, 1H), 0.60 - 0.51 (m, 1H), 0.38-0.32 (m, 1H) ppm. Compound 10' ret time 1.6 min, chiral purity >99% ee.

[0508] MS m / z (ESI): [M+H] + = 444.0

[0509] 1H NMR (400MHz, DMSO-d6) δ8.45(d,J=8.3Hz,1H),7.75(d,J=8.4Hz,1H),7.67(d,J=6.4Hz,2H), 7.59–7.53(m,2H),7.53–7.46(m,3H),7.04(dd,J=15.3,5.0Hz,1H),6.89(dd,J=15.4,1.4Hz ,1H), 4.92–4.77 (m,1H), 3.04 (s,3H), 1.95 (t,J=19.0Hz,3H), 1.82–1.69 (m,1H), 0.83–0.71 (m,1H), 0.68–0.62 (m,1H), 0.60–0.50 (m,1H), 0.40–0.31 (m,1H) ppm. Example 7 Synthesis of compounds 11k, 11 and 11'

[0510] Compound 11k

[0511] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0512] (E)-2-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-6-(1,1-Difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0513] Compounds 11 and 11'

[0514] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one and(R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0515] (S,E)-2-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-6-(1,1-Difluoroethyl)-4-phenylphthalazin-1(2H)-one and (R,E)-2-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-6-(1,1-Difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0516] first step

[0517] ethyl 4-acetyl-2-bromobenzoate

[0518] ethyl 4-acetyl-2-bromobenzoate

[0519] In a reaction vessel, 1-(3-bromo-4-iodophenyl)ethane-1-one 11a (15 g, 46.16 mmol, commercially available) was dissolved in ethanol (10 mL), and palladium on carbon (491.26 mg, 4.62 mmol, 10% wt) and triethylamine (14.01 g, 138.49 mmol) were added to replace carbon monoxide. The pressure was increased to 0.8 MPa, and the reaction was carried out at 50 °C for 16 hours. After returning to room temperature, the reaction solution was filtered through diatomaceous earth, and the filter cake was washed with ethanol (20 mL). The filtrate was collected, concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (eluent: system A) to give ethyl 4-acetyl-2-bromobenzoate 11b (9.8 g), with a yield of 78.31%.

[0520] MS m / z(ESI):[M+CH3CN+H] + =314 / 316

[0521] 1 H NMR(400MHz,DMSO-d6)δ8.20(d,J=1.6Hz,1H),8.02(dd,J=8.0,1.6Hz,1H),7.85 (d,J=8.0Hz,1H),4.36(q,J=7.1Hz,2H),2.62(s,3H),1.33(t,J=7.1Hz,3H)ppm.

[0522] Step 2

[0523] ethyl 2-bromo-4-(1,1-difluoroethyl)benzoate

[0524] ethyl 2-bromo-4-(1,1-difluoroethyl)benzoate

[0525] Ethyl 4-acetyl-2-bromobenzoate 11b (9.8 g, 36.15 mmol) was dissolved in diethylaminosulfur trifluoride (15 mL) and reacted at 50 °C for 16 hours. After returning to room temperature, the reaction solution was diluted with dichloromethane (20 mL) and added to a saturated sodium bicarbonate solution (50 mL). Extraction was performed with dichloromethane (10 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (eluent: system A) to give ethyl 2-bromo-4-(1,1-difluoroethyl)benzoate 11c (9.5 g), with a yield of 89.7%.

[0526] 1 H NMR (400 MHz, CDC13) δ 7.94 - 7.66 (m, 2H), 7.49 (d, J = 8.1 Hz, 1H), 4.42 (q, J = 7.1 Hz, 2H), 1.91 (t, J = 18.2 Hz, 3H), 1.41 (t, J = 7.1 Hz, 3H) ppm.

[0527] Third step

[0528] ethyl 4-(1,1-difluoroethyl)-2-(1-phenylvinyl)benzoate

[0529] ethyl 4-(1,1-difluoroethyl)-2-(1-phenylvinyl)benzoate

[0530] Ethyl 4-(1,1-difluoroethyl)-2-(1-phenylvinyl)benzoate 11d (5.0 g) was obtained in 92.7% yield. 1H NMR (400 MHz, CDC13) δ 7.94 - 7.66 (m, 2H), 7.49 (d, J = 8.1 Hz, 1H), 4.42 (q, J = 7.1 Hz, 2H), 1.91 (t, J = 18.2 Hz, 3H), 1.41 (t, J = 7.1 Hz, 3H) ppm.

[0531] MS m / z (ESI): [M+H] + = 317.2

[0532] Fourth step

[0533] ethyl 2-benzoyl-4-(1,1-difluoroethyl)benzoate

[0534] ethyl 2-benzoyl-4-(1,1-difluoroethyl)benzoate

[0535] Ethyl 2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetate 11a (1.2 g, 3.46 mmol) and potassium osmate (VI) dihydrate (291.18 mg, 790.29 μmol) were added to water (15 mL) under nitrogen atmosphere, sodium periodate (20.28 g, 94.83 mmol) was added, and the reaction was carried out at room temperature for 16 hours. Filtration was performed, and the filtrate was extracted with ethyl acetate (20 mL x 3), and the combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetate 11b (1.2 g) in a yield of 41.7%.

[0536] MS m / z (ESI): [M+H] + = 319.2

[0537] Fifth step

[0538] 6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0539] 6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0540] Ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetate 11a (1.2 g, 3.46 mmol) and potassium osmate (VI) dihydrate (291.18 mg, 790.29 μmol) were added to water (15 mL) under nitrogen atmosphere, sodium periodate (20.28 g, 94.83 mmol) was added, and the reaction was carried out at room temperature for 16 hours. Filtration was performed, and the filtrate was extracted with ethyl acetate (20 mL x 3), and the combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetate 11b (1.2 g) in a yield of 41.7%.

[0541] MS m / z (ESI): [M+H] + = 287.1

[0542] Sixth step

[0543] ethyl 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetate

[0544] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetate

[0545] To 6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one 11f (620 mg, 2.17 mmol) and 2-bromo-2-cyclopropylethyl acetate (717.52 mg, 3.47 mmol) in N,N- dimethylformamide (10 mL) was added cesium carbonate (1.13 g, 3.47 mmol) and the reaction was stirred at 80 °C for 16 h. The reaction was cooled to room temperature and water (15 mL) was added. The mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phase was concentrated under reduced pressure. The residue was separated by column chromatography on silica gel (eluent: system A) to give 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetic acid ethyl ester 11g (1.68 g, 97.4%). MS m / z (ESI): [M+H] + = 413.6

[0546] Seventh step

[0547] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0548] 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0549] To 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetic acid ethyl ester 11g (1.5 g, 3.64 mmol) in ethanol (10 mL) was added sodium borohydride (165.12 mg, 4.36 mmol) at 0 °C. The reaction was stirred at room temperature for 12 h. Saturated ammonium chloride solution (20 mL) was added. The mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phase was concentrated under reduced pressure. The residue was separated by column chromatography on silica gel (eluent: system A) to give 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one 11h (930 mg, 69.0% yield).

[0550] MS m / z (ESI): [M+H] + = 371.3

[0551] Eighth step

[0552] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetaldehyde

[0553] 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetaldehyde

[0554] Oxalyl chloride (2.55 g, 20.09 mmol) was added to dichloromethane (30 mL) and cooled to -70 °C. A solution of dimethyl sulfoxide (1.96 g, 25.11 mmol) in dichloromethane (10 mL) was added dropwise slowly and the reaction was kept at -70 °C for 30 min. A solution of 2-(1-cyclopropyl-2-hydroxyethyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one 11h (930 mg, 2.51 mmol) in dichloromethane (10 mL) was added and the reaction was kept at -70 °C for 1 h. A solution of triethylamine (4.57 g, 45.20 mmol) in dichloromethane (10 mL) was added and the reaction was kept at room temperature for 1 h. The reaction was added to saturated ammonium chloride solution (50 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetaldehyde 11i (900 mg) which was used directly in the next step.

[0555] MS m / z (ESI): [M+H] + = 369.2

[0556] Ninth step

[0557] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0558] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0559] ((methylsulfonyl)methyl)phosphonic acid diethyl ester 1j (843.69 mg, 3.66 mmol) was dissolved in tetrahydrofuran (20 mL) under nitrogen atmosphere, cooled to -70 °C, n-butyllithium solution in n-hexane (1.6 mol / L, 1.99 mL) was added, the reaction was continued at -70 °C for 1 hour, 2-cyclopropyl-2-(6-(1,1-difluoroethyl)-1-oxo-4-phenylphthalazin-2(1H)-yl)acetaldehyde 11i (900 mg, 2.44 mmol) in tetrahydrofuran (20 mL) was added, the reaction was continued at -70 °C for 1 hour, saturated ammonium chloride solution (20 mL) was added to quench, the temperature was allowed to recover to room temperature, ethyl acetate (30 mL x 3) was used to extract, the combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, the obtained residue was purified by preparative liquid phase separation (Waters 3767 / Qda, column: XBridge C 18 , 19*250 mm, 10 μm; Mobile Phase A: 10 mmol / L ammonium bicarbonate aqueous solution, mobile phase B: acetonitrile; flow rate: 20 mL / min), to obtain (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one 11k (144.39 mg), yield 13.3%.

[0560] MS m / z (ESI): [M+H] + = 445.2

[0561] 1 H NMR (400 MHz, DMSO-d6) δ 8.51 (d, J = 8.4 Hz, 1H), 8.10 (dd, J = 8.3, 1.3 Hz, 1H), 7.84 (s, 1H), 7.71 - 7.65 (m, 2H), 7.64 - 7.57 (m, 3H), 7.04 - 6.93 (m, 2H), 5.08 (dd, J = 9.9, 3.6 Hz, 1H), 3.02 (s, 3H), 2.00 (t, J = 19.1 Hz, 3H), 1.63 - 1.51 (m, 1H), 0.78 - 0.64 (m, 2H), 0.59 - 0.48 (m, 1H), 0.44 - 0.34 (m, 1H) ppm.

[0562] Tenth step

[0563] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0564] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one

[0565] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-(1,1-difluoroethyl)-4-phenylphthalazin-1(2H)-one 11k (136 mg) was purified by chiral prep separation (Chiral column: 250*25 mm, 10 μm, flow rate 100 mL / min, gradient: 45% MeOH: 55% CO2) to give the longer retention time single enantiomer compound 11 (32 mg) in 23.5% yield and the shorter retention time single enantiomer compound 11' (35 mg) in 25.7% yield.

[0566] Chiral analysis method:

[0567] Chromatography column: 100*3 mm, 3 μm;

[0568] Flow rate: 2.0 mL / min;

[0569] Gradient: 65% MeOH (with 0.1% diethylamine): 35% CO2.

[0570] Compound 11 retention time 2.78 min, chiral purity >99% ee.

[0571] MS m / z (ESI): [M+H] + = 445.2

[0572] 1H NMR (400MHz, DMSO) δ8.51(d,J=8.3Hz,1H),8.10(dd,J=8.4,1.6Hz,1H),7.84(s,1H),7.71–7.65(m,2H),7.65–7.58(m,3H),7.04–6.93(m,2H),5 .08(dd,J=9.9,3.7Hz,1H),3.02(s,3H),2.00(t,J=19.1Hz,3H),1.63–1 .48(m,1H),0.78–0.63(m,2H),0.58–0.49(m,1H),0.45–0.35(m,1H)ppm.

[0573] Compound 11' has a retention time of 1.37 minutes and a chiral purity of >99% ee.

[0574] MS m / z(ESI):[M+H] + =445.2

[0575] 1 H NMR (400MHz, DMSO) δ8.51(d,J=8.4Hz,1H),8.10(dd,J=8.4,1.6Hz,1H),7.84(s,1H),7.70–7.65(m,2H),7.64–7.58(m,3H),7.04–6.92(m,2H),5 .08(dd,J=9.9,3.7Hz,1H),3.02(s,3H),2.00(t,J=19.1Hz,3H),1.62–1 .51(m,1H),0.78–0.64(m,2H),0.58–0.49(m,1H),0.44–0.35(m,1H)ppm.

[0576] Example 8 Synthesis of compounds 12k, 12 and 12'

[0577] Compound 12k

[0578] (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0579] (E)-2-(1-Cyclopropyl-3-(Methanesulfonyl)allyl)-4-Phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0580] Compounds 12 and 12'

[0581] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0582] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0583] First step

[0584] 4-bromo-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0585] 4-bromo-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0586] 2,9-dihydro-1H-pyrido[3,4-b]indol-1-one 12a (2.5 g, 13.57 mmol, prepared according to published patent WO2001009129) was dissolved in acetonitrile (50 mL), N-bromosuccinimide (2.42 g, 13.57 mmol) and acetic acid (0.1 mL) were added and the reaction was allowed to proceed at room temperature for 1 hour. The reaction was concentrated under reduced pressure and the residue was separated by column chromatography on silica gel (eluent: system B) to give 4-bromo-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one 12b (832 mg) in 23.3% yield.

[0587] MS m / z (ESI): [M+H] + = 263.0 / 265.0

[0588] Second step

[0589] 4-bromo-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0590] 4-bromo-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0591] Under nitrogen atmosphere, 4-bromo-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one 12b (832 mg, 3.16 mmol) was dissolved in a mixed solvent of 1,4-dioxane (10 mL) and water (1 mL), and benzeneboronic acid (771.18 mg, 6.32 mmol), bis(diphenylphosphino)ethane palladium dichloride (117.29 mg, 158.12 μmol) and potassium carbonate (1.31 g, 9.49 mmol) were added, and the mixture was reacted at 100 °C for 2 hours. After being cooled to room temperature, ethyl acetate (30 mL) was added, and the mixture was washed with saturated sodium chloride solution (10 mL). The organic phase was concentrated under reduced pressure, and the obtained residue was separated by silica gel column chromatography (eluent: B system) and then by preparative liquid chromatography (Isolera-Biotage column: Agela C 18 40g; mobile phase A: 0.1% trifluoroacetic acid aqueous solution, mobile phase B: acetonitrile; flow rate: 20 mL / min;), to give 4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one 12c (205 mg) in a yield of 24.9%. MS m / z (ESI): [M+H] + = 261.0

[0592] 1 H NMR (400 MHz, DMSO-d6) δ 12.16 (s, 1H), 11.58 (d, J = 5.6 Hz, 1H), 7.56 - 7.52 (m, 4H), 7.51 - 7.44 (m, 2H), 7.40 - 7.31 (m, 2H), 7.01 - 6.89 (m, 2H) ppm.

[0593] Third step

[0594] ethyl 2-cyclopropyl-2-(1-oxo-4-phenyl-1,9-dihydro-2H-pyrido[3,4-b]indol-2-yl)acetate

[0595] 2-cyclopropyl-2-(1-oxo-4-phenyl-1,9-dihydro-2H-pyrido[3,4-b]indol-2-yl)acetate

[0596] To a solution of 4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one 12c (150 mg, 576.3 μmol) in N,N-dimethylformamide (3 mL) was added cesium carbonate (375.6 mg, 1.15 mmol), 2-bromo-2-cyclopropylethyl acetate (119.4 mg, 576.3 μmol) and the reaction mixture was heated at 80 °C for 16 h. The reaction mixture was cooled to room temperature and purified by preparative liquid chromatography (Isolera-Biotage chromatography column: Agela C18 10 μm 30x150 mm; eluent A: 0.1% trifluoroacetic acid in water, eluent B: acetonitrile; flow rate: 20 mL / min) to give 2-cyclopropyl-2-(1-oxo-4-phenyl-1,9-dihydro-2H-pyrido[3,4-b]indol-2-yl)ethyl acetate 12d (150 mg) in 67.4% yield. 18 12g; mobile phase A: 0.1% trifluoroacetic acid in water, mobile phase B: acetonitrile; flow rate: 20 mL / min), in 67.4% yield.

[0597] MS m / z (ESI): [M+H] + = 387.3

[0598] 1 H NMR (400 MHz, DMSO-d6) δ 12.23 (s, 1H), 7.61 - 7.49 (m, 6H), 7.46 - 7.31 (m, 3H), 7.04 - 6.96 (m, 1H), 4.56 - 4.45 (m, 1H), 4.26 - 4.08 (m, 2H), 1.84 - 1.66 (m, 1H), 1.20 (t, J = 7.1 Hz, 3H), 0.88 - 0.75 (m, 1H), 0.72 - 0.62 (m, 1H), 0.61 - 0.49 (m, 1H), 0.45 - 0.30 (m, 1H) ppm.

[0599] Fourth step

[0600] 2-(1-cyclopropyl-2-hydroxyethyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0601] 2-(1-cyclopropyl-2-hydroxyethyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0602] Ethyl 2-cyclopropyl-2-(l-oxo-4-phenyl-l,9-dihydro-2H-pyrido[3,4-b]indol-2- yl)acetate 12d (140 mg, 362.28 μmol) was dissolved in ethanol (5 mL), and sodium borohydride (68.53 mg, 1.81 mmol) was added. The reaction was stirred at room temperature for 2 hours. The reaction was concentrated under reduced pressure, and the residue was separated by column chromatography on silica gel (eluent: system B) to give 2-(l-cyclopropyl-2-hydroxyethyl)-4-phenyl-2,9-dihydro-lH- pyrido[3,4-b]indol-l-one 12e (54 mg) in 43.3% yield.

[0603] MS m / z (ESI): [M+H] + = 345.0

[0604] 1 H NMR (400 MHz, DMSO) δ 12.11 (s, 1H), 7.63 - 7.59 (m, 2H), 7.57 - 7.47 (m, 5H), 7.40 (s, 1H), 7.38 - 7.34 (m, 1H), 7.00 (t, J = 7.1 Hz, 1H), 4.94 (t, J = 5.5 Hz, 1H), 4.44 - 4.21 (m, 1H), 4.07 - 3.74 (m, 2H), 1.51 - 1.41 (m, 1H), 0.75 - 0.63 (m, 1H), 0.59 - 0.49 (m, 1H), 0.49 - 0.42 (m, 1H), 0.30 - 0.20 (m, 1H) ppm.

[0605] Fifth step

[0606] 2-cyclopropyl-2-(l-oxo-4-phenyl-l,9-dihydro-2H-pyrido[3,4-b]indol-2-yl)acetaldehyde

[0607] 2-cyclopropyl-2-(l-oxo-4-phenyl-l,9-dihydro-2H-pyrido[3,4-b]indol-2-yl)acetaldehyde

[0608] Oxalyl chloride (159.21 mg, 1.25 mmol) was added to dichloromethane (5 mL) under nitrogen atmosphere, cooled to -70 °C, a solution of dimethyl sulfoxide (122.50 mg, 1.57 mmol) in dichloromethane (3 mL) was added dropwise, kept at -70 °C for 30 min, a solution of 2-(l-cyclopropyl-2-hydroxyethyl)-4-phenyl-2,9-dihydro-lH-pyrido[3,4-b]indol-l-one 12e (54 mg, 156.79 μmol) in dichloromethane (3 mL) was added dropwise, kept at -70 °C for 1 h, a solution of triethylamine (237.99 mg, 2.35 mmol) in dichloromethane (3 mL) was added dropwise, warmed to room temperature for 1 h, the reaction was quenched with saturated ammonium chloride solution (5 mL), extracted with dichloromethane (10 mL x 2), the combined organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure to give 2-cyclopropyl-2-(l-oxo-4-phenyl-l,9-dihydro-2H-pyrido[3,4-b]indol-2-yl)acetaldehyde 12f (53 mg), which was used directly in the next step.

[0609] MS m / z (ESI): [M+H] + = 343.2

[0610] Sixth step

[0611] (E)-2-(l-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-lH-pyrido[3,4-b]indol-l-one

[0612] (E)-2-(l-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-lH-pyrido[3,4-b]indol-l-one

[0613] ((Methylsulfonyl)methyl)phosphonic acid diethyl ester 1j (67.24 mg, 292.06 μmol) was dissolved in tetrahydrofuran (5 mL), cooled to -70 °C, n-butyllithium in n-hexane (1.6 M, 127.78 μL) was added, -70 °C for 1 h, a solution of 2-cyclopropyl-2-(l-oxo-4-phenyl-l,9-dihydro-2H-pyrido[3,4-b]indol-2-yl)acetaldehyde 12f (50 mg, 146.03 μmol) in tetrahydrofuran (5 mL) was added, -70 °C for 1 h, quenched with saturated ammonium chloride solution (3 mL), warmed to room temperature, extracted with ethyl acetate (5 mL x 3), the combined organic phase was washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, the obtained residue was purified by preparative liquid phase separation (Waters 3767 / Qda column: Sunfire C18 OBD Prep 10 x 150 mm 5 μm; mobile phase: acetonitrile: water = 60:40; flow rate: 10 mL / min; detection: 254 nm), to give (E)-2-(l-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-lH-pyrido[3,4-b]indol-l-one (20 mg, 35.62% yield).18 , 19*250 mm, 10 μm; mobile phase A: 0.1% formic acid in water, B: acetonitrile; flow rate: 20 mL / min); to give (E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one 12k (6.5 mg) in 10.6% yield.

[0614] MS m / z (ESI): [M+CH3CN+H] + = 460.1

[0615] 1 H NMR (400 MHz, MeOH-d4) δ 7.64 - 7.58 (m, 3H), 7.57 - 7.48 (m, 3H), 7.45 - 7.38 (m, 2H), 7.29 (s, 1H), 7.15 (dd, J = 15.5, 4.7 Hz, 1H), 7.00 (t, J = 7.4 Hz, 1H), 6.80 (dd, J = 15.4, 2.0 Hz, 1H), 5.13 - 5.03 (m, 1H), 3.01 (s, 3H), 1.73 - 1.60 (m, 1H), 1.37 - 1.27 (m, 1H), 0.97 - 0.88 (m, 1H), 0.72 - 0.62 (m, 2H), 0.57 - 0.46 (m, 1H) ppm.

[0616] Seventh step

[0617] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0618] (S,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one and (R,E)-2-(1-cyclopropyl-3-(methylsulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one

[0619] (E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-4-phenyl-2,9-dihydro-1H-pyrido[3,4-b]indol-1-one 12kJ (400mg) was purified by chiral preparation (chiral column: The flow rate was 140 mL / min, and the gradient was 60% MeOH and 40% isopropanol. The results showed that a monomorphic compound 12 (125 mg, yield 31.3%) with a longer retention time and a monomorphic compound 12' (101 mg, yield 25.3%) with a shorter retention time were obtained.

[0620] Chiral analysis method

[0621] Chromatographic column: 100*3.0mm, 3μm;

[0622] Flow rate: 1.5 mL / min;

[0623] Gradient: 35% isopropanol (containing 0.1% diisopropylamine): 65% CO2.

[0624] Compound 12 has a retention time of 4.1 minutes and a chiral purity of >99% ee.

[0625] 1 H NMR (400MHz, DMSO-d) 6 )δ12.25(s,1H),7.64–7.59(m,2H),7.59–7.53(m,3H),7.52–7.46(m,1H),7.42 –7.35(m,3H),7.06(dd,J=15.3,5.2Hz,1H),7.01(t,J=8.0Hz,1H),6.85(dd,J= 15.3,1.5Hz,1H),5.00(dd,J=8.9,5.1Hz,1H),3.05(s,3H),1.83–1.71(m,1H), 0.84–0.73(m,1H),0.71–0.63(m,1H),0.61–0.52(m,1H),0.44–0.33(m,1H)ppm.

[0626] MS m / z(ESI):[M+H] + =419.0

[0627] Compound 12' has a retention time of 2.4 minutes and a chiral purity of 98.6% ee.

[0628] MS m / z(ESI):[M+H] + =419.3

[0629] 1H NMR (400MHz, DMSO-d) 6 )δ12.25(s,1H),7.65–7.60(m,2H),7.59–7.53(m,3H),7.52–7.46(m,1H),7.43 –7.34(m,3H),7.06(dd,J=15.3,5.2Hz,1H),7.01(t,J=7.6Hz,1H),6.85(dd,J= 15.3,1.5Hz,1H),5.00(dd,J=8.7,5.2Hz,1H),3.05(s,3H),1.83–1.71(m,1H), 0.82–0.74(m,1H),0.72–0.62(m,1H),0.60–0.53(m,1H),0.42–0.34(m,1H)ppm.

[0630] Example 9 Synthesis of Compound 13k

[0631] Compound 13k

[0632] (E)-8-cyclohexyl-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0633] (E)-8-cyclohexyl-6-(1-cyclopropyl-3-(methanesulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0634] first step

[0635] (S)-8-cyclohexyl-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0636] (S)-8-cyclohexyl-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0637] Iodocyclohexane (1.40 g, 6.68 mmol) and (S)-8-bromo-6-(1-cyclopropyl-2- hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 1g (500 mg, 1.34 mmol) were dissolved in N,N-dimethylacetamide (6 mL) under nitrogen atmosphere, zinc powder (524.27 mg, 8.02 mmol), nickel dichloride dimethoxyethane (58.72 mg, 267.25 μmol) and 2-picolinamidine hydrochloride (42.12 mg, 267.25 μmol) were added, and the reaction was carried out at 100 °C for 4 hours. After being cooled to room temperature, the reaction solution was diluted with water (30 mL) and extracted with dichloromethane (30 mL x 3). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain (S)-8-cyclohexyl-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 13a (81 mg) in a yield of 16.1%.

[0638] MS m / z (ESI): [M+H] + = 378.3

[0639] Second step

[0640] 2-(8-cyclohexyl-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde

[0641] 2-(8-cyclohexyl-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde

[0642] Oxalyl chloride (242.13 mg, 1.91 mmol) was dissolved in dichloromethane (3 mL) and cooled to -70 °C, a solution of dimethyl sulfoxide (186.31 mg, 2.38 mmol) in dichloromethane (3 mL) was slowly added, and the reaction was maintained at -70 °C for 30 min. Then a solution of (S)-8-cyclohexyl-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 13a (90 mg, 238.46 μmol) in dichloromethane (3 mL) was added, and the reaction was maintained at -70 °C for 1 h. Then triethylamine (361.94 mg, 3.58 mmol) was added dropwise. The reaction was allowed to reach room temperature and was maintained for 1 h. The reaction was diluted with water (20 mL) and extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 2-(8-cyclohexyl-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde 13b (81 mg) in 90.5% yield.

[0643] MS m / z (ESI): [M+H] + = 376.2

[0644] Third step

[0645] (E)-8-cyclohexyl-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0646] (E)-8-cyclohexyl-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0647] ((methylsulfonyl)methyl)phosphonic acid diethyl ester 1j (82.79 mg, 359.61 μmol) was dissolved in tetrahydrofuran (2 mL) under nitrogen atmosphere, cooled to -78 °C, n-butyllithium in n-hexane (1.6 mol / L, 195 μL) was added, and the reaction was continued at -78 °C for 30 minutes. A solution of 2-(8-cyclohexyl-2-(1,1-difluoroethyl)-5-oxopyridino[4,3-d]pyrimidin-6(5H)-yl)-2- cyclopropylacetaldehyde 13b (90 mg, 239.74 μmol) in tetrahydrofuran (2 mL) was added, and the reaction was continued at -78 °C for 20 minutes. The reaction was quenched by adding saturated ammonium chloride solution (5 mL), and the temperature was allowed to rise to room temperature. The organic phase was extracted with dichloromethane (5 mL x 3), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography (Waters 3767 / Qda column: SunFire Sunfire C18, 19*250mm, 10μm; mobile phase A: 0.1% formic acid in water, mobile phase B: acetonitrile; flow rate: 20 mL / min) to give (E)-8-cyclohexyl-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 13k (13.35 mg) in 12.3% yield.

[0648] MS m / z (ESI): [M+H] + = 452.3

[0649] 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.60 (s, 1H), 7.90 (s, 1H), 7.00 (dd, J = 15.3, 4.8 Hz, 1H), 6.86 (dd, J = 15.3, 1.6 Hz, 1H), 4.75 (dd, J = 10.0, 4.7 Hz, 1H), 3.16 (t, J = 11.7 Hz, 1H), 3.01 (s, 3H), 2.10 (t, J = 19.0 Hz, 3H), 1.91 - 1.80 (m, 4H), 1.79 - 1.72 (m, 2H), 1.62 - 1.52 (m, 2H), 1.48 - 1.37 (m, 2H), 1.28 - 1.23 (m, 1H), 0.80 (s, 1H), 0.70 - 0.64 (m, 1H), 0.54 (s, 1H), 0.32 (dd, J = 9.6, 5.2 Hz, 1H) ppm.

[0650] Synthesis of compound 14k of Example 10

[0651] Compound 14k

[0652] (E)-8-cyclopentyl-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0653] (E)-8-cyclopentyl-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0654] Compound 14k was synthesized according to the procedure for Compound 13k in Reference Example 9, by replacing iodocyclohexane with iodocyclopentane. MS m / z (ESI): [M+H] + = 438.3

[0655] 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.60 (s, 1H), 7.94 (s, 1H), 7.00 (dd, J = 15.4, 4.7 Hz, 1H), 6.88 (dd, J = 15.4, 1.5 Hz, 1H), 4.76 (dd, J = 8.8, 4.8 Hz, 1H), 3.02 (s, 3H), 2.15 - 2.02 (m, 5H), 1.87 - 1.62 (m, 8H), 0.84 - 0.76 (m, 1H), 0.70 - 0.63 (m, 1H), 0.58 - 0.51 (m, 1H), 0.36 - 0.30 (m, 1H) ppm.

[0656] Synthesis of compound 15k in Example 11

[0657] Compound 15k

[0658] (E)-8-(cyclohex-1-en-1-yl)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0659] (E)-8-(cyclohex-1-en-1-yl)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0660] First step

[0661] 2-(8-bromo-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde

[0662] 2-(8-bromo-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde

[0663] Oxalyl chloride (814.13 mg, 6.41 mmol) was dissolved in dichloromethane (6 mL) and cooled to -70 °C. A solution of dimethyl sulfoxide (626.41 mg, 8.02 mmol) in dichloromethane (6 mL) was added dropwise slowly, and the reaction was maintained at -70 °C for 30 min. A solution of 8-bromo-6-(1-cyclopropyl-2-hydroxyethyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 1g (300 mg, 801.75 μmol) in dichloromethane (6 mL) was added, and the reaction was maintained at -70 °C for 1 h. Triethylamine (1.22 g, 12.03 mmol) was added dropwise, and the reaction was maintained at room temperature for 1 h. The reaction was quenched with saturated ammonium chloride solution (5 mL), diluted with water (10 mL), and extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 2-(8-bromo-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde 15a (290 mg) in 97.2% yield.

[0664] MS m / z (ESI): [M+H] + = 372.1 / 374.1

[0665] Second step

[0666] 8-bromo-6-(cyclopropyl(1,3-dioxolan-2-yl)methyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0667] 8-bromo-6-(cyclopropyl(1,3-dioxolan-2-yl)methyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0668] To a solution of 2-(8-bromo-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2- cyclopropylacetaldehyde 15a (500 mg, 1.34 mmol) in toluene (7 mL) was added ethylene glycol (333.56 mg, 5.37 mmol) and p-toluenesulfonic acid (51.11 mg, 268.70 μmol) and the reaction was stirred at 120 °C for 12 h. The reaction was cooled to room temperature and concentrated under reduced pressure. The residue was diluted with water (15 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: A system) to give 8-bromo-6-(cyclopropyl(1,3-dioxolan-2-yl)methyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 15b (116 mg) in 20.7% yield.

[0669] MS m / z (ESI): [M+H] + = 416.0 / 418.0

[0670] Third step

[0671] 8-(cyclohex-1-en-1-yl)-6-(cyclopropyl(1,3-dioxolan-2-yl)methyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0672] 8-(cyclohex-1-en-1-yl)-6-(cyclopropyl(1,3-dioxolan-2-yl)methyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0673] Under nitrogen atmosphere, 8-bromo-6-(cyclopropyl(l,3-dioxolan-2-yl)methyl)-2-(l,l- difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 15b (106 mg, 254.68 pmol) and cyclohexen- 1- boronic acid pinacol ester (63.60 mg, 305.61 pmol) were dissolved in a mixed solvent of tetrahydrofuran (3 mL) and water (0.6 mL), potassium phosphate (135.15 mg, 636.69 pmol) and CataCXium A Pd G3 (CAS No.: 1651823-59-4, 18.57 mg, 25.47 pmol) were added, and the reaction was carried out at 100 °C for 2 hours. After being restored to room temperature, the reaction solution was diluted with water (10 mL), extracted with ethyl acetate (10 mL x 3), and the organic phase was combined. The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained residue was separated by silica gel column chromatography (eluent: A system) to obtain 8-(cyclohex-1-en-1-yl)-6-(cyclopropyl(l,3-dioxolan-2-yl)methyl)-2-(l,l- difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 15c (86 mg), with a yield of 83.7%. MS m / z (ESI): [M+H] + = 418.5

[0674] Fourth step

[0675] 2-(8-(cyclohex-1-en-1-yl)-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde

[0676] 2-(8-(cyclohex-1-en-1-yl)-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde

[0677] 8-(cyclohexyl-1-en-1-yl)-6-(cyclopropyl(1,3-dioxopentane-2-yl)methyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 15c (70 mg, 167.69 μmol) was dissolved in tetrahydrofuran (3 mL), and dilute hydrochloric acid solution (3 mol / L, 0.3 mL) was added. The mixture was reacted at 70 °C for 12 hours. The reaction mixture was brought to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (eluent: system A) to give 15d (8 mg) of 2-(8-(cyclohexyl-1-en-1-yl)-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde, yield 12.8%. MS m / z (ESI): [M+H] + =374.1

[0678] Step 5

[0679] (E)-8-(cyclohex-1-en-1-yl)-6-(1-cyclopropyl-3-(methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0680] (E)-8-(cyclohex-1-en-1-yl)-6-(1-cyclopropyl-3-(methanesulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one

[0681] Dissolve 100 mg (7.40 μmol) of diethyl ((methanesulfonyl)methyl)phosphate in 1 mL of tetrahydrofuran. Cool to -78 °C and add 13.4 μL of n-butyllithium in hexane. Maintain the reaction temperature at -78 °C for 30 minutes. Then add 150 mg (8 mg, 21.42 μmol) of 2-(8-(cyclohex-1-en-1-yl)-2-(1,1-difluoroethyl)-5-oxopyrido[4,3-d]pyrimidin-6(5H)-yl)-2-cyclopropylacetaldehyde in 1 mL of tetrahydrofuran. Maintain the reaction temperature at -78 °C for another 20 minutes. Quenching was performed with saturated ammonium chloride solution (5 mL), and the mixture was allowed to return to room temperature. Extraction was then performed with dichloromethane (7 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was then purified by preparative liquid chromatography (Waters 3767 / QDA). Column: SunFire C 18,19*250 mm, 10 pm; mobile phase A: 0.1% formic acid in water, mobile phase B: acetonitrile; flow rate: 20 mL / min) to give (E)-8-(cyclohex-1-en-1-yl)-6-(1-cyclopropyl-3- (methylsulfonyl)allyl)-2-(1,1-difluoroethyl)pyrido[4,3-d]pyrimidin-5(6H)-one 15k (0.83 mg), yield 8.6%.

[0682] MS m / z (ESI): [M+H] + = 450.2

[0683] 1 H NMR (400 MHz, DMSO) δ 9.60 (s, 1H), 7.99 (s, 1H), 7.05 - 6.86 (m, 2H), 5.99 (s, 1H), 4.78 - 4.71 (m, 1H), 3.02 (s, 3H), 2.55 - 2.53 (m, 2H), 2.23 - 2.17 (m, 2H), 2.08 (t, J = 19.0 Hz, 3H), 1.80 - 1.62 (m, 5H), 0.84 - 0.75 (m, 1H), 0.73 - 0.62 (m, 1H), 0.60 - 0.51 (m, 1H), 0.40 - 0.29 (m, 1H) ppm.

[0684] Biological evaluation

[0685] Test Example 1, Inhibition of SW48 cell proliferation by compounds of the application

[0686] The following method was used to determine the effect of compounds of the application on SW48 cell proliferation. SW48 cells (MSI-H cells) were purchased from the ATCC cell bank in the USA and cultured in Leibovitz's L-15 (Gibco, Cat. No. 11415064) medium containing 10% fetal bovine serum, 100 U of penicillin and 100 pg / mL of streptomycin. Cell viability was determined by Luminescent Cell Viability Assay kit (Promega, Cat. No. G7573).

[0687] The experimental method is operated according to the steps of the kit instruction, and the brief description is as follows: the test compound is first dissolved in DMSO to prepare a 10 mM stock solution, and then diluted with the above-mentioned culture medium to prepare a test sample, and the final concentration of the compound ranges from 10000 nM to 1.52 nM. Cells in the logarithmic growth phase are inoculated into a 96-well cell culture plate at a density of 500 cells per well, incubated in a 37°C air incubator overnight, and then continue to incubate for 120 hours after adding the test compound. After the end of the culture, 40 uL of CellTiter-Glo detection solution is added to each well, shaken for 5 minutes, and then placed for 10 minutes, and then the luminescence value of each well of the sample is read on the enzyme label instrument using the Luminescence mode. The percentage inhibition rate of the compound at each concentration point is calculated by comparing the value with the control group (0.1% DMSO), and then the compound concentration logarithm-inhibition rate is nonlinearly regressed in the GraphPad Prism 9 software to obtain the IC 50 value of the compound for inhibiting cell proliferation.

[0688] The IC 50 value of the compound of the present application is represented by AA, A, respectively, as follows:

[0689] AA: IC 50 ≤ 10 nM

[0690] A: 10 < IC 50 ≤ 100 nM

[0691] Conclusion: The compound of the present application has excellent inhibitory effect on the proliferation of SW48 cells.

[0692] Test Example 2, Determination of the Inhibition of the Proliferation of RL95-2 Cells by the Compound of the Present Invention

[0693] The following method is used to determine the effect of the compound of the present application on the proliferation of RL95-2 cells. RL95-2 cells (MSI-H cells) are purchased from the Cell Resource Center of Shanghai Life Sciences Research Institute, Chinese Academy of Sciences, and cultured in DMEM / F12 medium (Gibco, catalog number A4192001) containing 10% fetal bovine serum, 100 U of penicillin and 100 μg / mL of streptomycin. Cell viability is determined by Luminescent Cell Viability Assay kit (Promega, catalog number G7573).

[0694] The experimental method is operated according to the steps of the kit instruction, and the brief description is as follows: the test compound is first dissolved in DMSO to prepare a 10 mM stock solution, then diluted with the above-mentioned culture medium to prepare a test sample, and the final concentration of the compound ranges from 10000 nM to 1.52 nM. Cells in the logarithmic growth phase are inoculated into a 96-well cell culture plate at a density of 500 cells per well, incubated in a 37°C, 5% CO2 incubator overnight, and then incubated for 120 hours after the addition of the test compound. After the end of the culture, 50 uL of CellTiter-Glo detection solution is added to each well, shaken for 5 minutes, and then incubated for 10 minutes. Then, the luminescence value of each well of the sample is read on a microplate reader using the Luminescence mode. The percentage inhibition of the compound at each concentration point is calculated by comparing the values with the control group (0.1% DMSO), and then the compound concentration logarithm-inhibition rate is analyzed by non-linear regression analysis in GraphPad Prism 9 software to obtain the IC 50 value of the compound for inhibiting cell proliferation.

[0695] The IC 50 value of the compound of the present application is represented by AA, A, respectively, as follows:

[0696] AA: IC 50 ≤ 10 nM

[0697] A: 10 nM < IC 50 < 100 nM

[0698] Conclusion: The compound of the present application has a good inhibitory effect on the IC 50 < 100 nM of RL95-2 cell proliferation.

[0699] Test Example 3, determination of the inhibitory effect of the compound of the present application on HCT116 cell proliferation

[0700] The following method is used to determine the effect of the compound of the present application on HCT116 cell proliferation. HCT116 cells are purchased from the Cell Resource Center of Shanghai Life Sciences Research Institute, Chinese Academy of Sciences, and cultured in McCoy's 5a medium containing 10% fetal bovine serum, 100 U of penicillin and 100 μg / mL of streptomycin. Cell viability is determined by Luminescent Cell Viability Assay kit (Promega, item number G7573).

[0701] The experimental method is operated according to the steps of the kit instruction, and the brief description is as follows: the test compound is first dissolved in DMSO to prepare a 10 mM stock solution, then diluted with the above-mentioned culture medium to prepare a test sample, and the final concentration of the compound is in the range of 10000 nM-1.52 nM. Cells in the logarithmic growth phase are inoculated into a 96-well cell culture plate at a density of 300 cells per well, cultured in a 37℃, 5%CO2 incubator overnight, and then continue to be cultured for 120 hours after adding the test compound. After the end of the culture, 50uL of CellTiter-Glo detection solution is added to each well, shaken for 5 minutes, and then placed for 10 minutes, and then the luminescence value of each well of the sample is read on the enzyme label instrument using the Luminescence mode. The percentage inhibition rate of the compound at each concentration point is calculated by comparing the value with the control group (0.1% DMSO), and then the compound concentration-log-inhibition rate is nonlinearly regressed in the GraphPad Prism 9 software to obtain the IC 50 value of the compound for inhibiting cell proliferation.

[0702] The IC 50 value of the compound of the present application is represented by AA, A, respectively, as follows:

[0703] AA: IC 50 ≤10nM

[0704] A: 10nM < IC 50 <100nM

[0705] Conclusion: The compound of the present application has a good inhibitory effect on the IC 50 <100nM of HCT116 cell proliferation. Test Example 4, mouse pharmacokinetic test of the compound of the present application

[0706] 1. Experimental purpose

[0707] ICR mice are used as test animals, and the LC / MS / MS method is used to determine the drug concentration in the plasma at different times after intragastrically administering the compound of the present application, to study the pharmacokinetic characteristics of the compound of the present application in mice.

[0708] 2. Experimental scheme

[0709] 2.1 Experimental drugs and animals

[0710] The compound of the present application

[0711] ICR mice, male, 29.5-36.8g, purchased from Vantianlihua Experimental Animal Technology Co., Ltd.

[0712] 2.2 Drug preparation

[0713] An appropriate amount of compound was weighed, and an appropriate amount of DMA, Kolliphor ELP and saline was added in turn, and ultrasonic vortex was mixed to prepare a 1 mg / mL administration preparation. Among them, DMA, Kolliphor ELP and saline = 5:5:90 (v:v:v).

[0714] 2.3 Administration

[0715] ICR mice, each test compound injection group (9 in each group), after overnight fasting, oral administration (PO, compound administration dose is 10 mg / kg, administration volume is 10 mL / kg), and food after 4 hours.

[0716] 3. Operation

[0717] About 0.1 mL of blood was taken from the orbit before administration and at 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 10 hours and 24 hours after administration. The whole blood sample was placed in an anticoagulant tube containing EDTA-K2. After blood sample collection, it was placed on ice and centrifuged to separate plasma (centrifugation conditions: 1500g, 10 minutes). The collected plasma was stored at -40 to -20℃ before analysis.

[0718] LC-MS / MS was used to determine the content of the test compound in the plasma of mice after oral administration.

[0719] 4. Pharmacokinetic parameter results

[0720] The pharmacokinetic parameters of the compound of the present application are shown in the following table.

[0721] Pharmacokinetic parameters of the compound of the present application in mice

[0722] Conclusion: The blood concentration and area under the curve of the compound of the present application are both high, and it has good pharmacokinetic properties.

[0723] BALB / c nude mice were subcutaneously inoculated with RL95-2 human endometrial cancer cells to establish a subcutaneous xenotransplant tumor model. The test was divided into test substance RO7589831 treatment group, test compound 10 treatment group and solvent control group, 6 in each group. Oral gavage administration for 20 days. The efficacy was evaluated according to the tumor growth, and the safety was evaluated according to the animal body weight change and death.

[0724] BALB / c nude mice, female, 6-7 weeks, purchased from Hangzhou Qizhen Experimental Animal Technology Co., Ltd. The animal experiment was approved by the experimental animal welfare ethics committee. The animals were fed in the experimental environment for at least 7 days before the experiment began. The experimental animals were fed in a constant temperature and humidity intelligent independent ventilation cage (IVC), the temperature of the feeding room was 20-26℃, the humidity was 40-70%, and the animals could freely take food and water. The experimental animals were labeled by ear tag method.

[0725] RL95-2 human endometrial cancer cells were cultured in DMEM and F12 culture medium containing 10% fetal bovine serum and 1% penicillin-streptomycin-bothymicin B solution. Exponentially growing RL95-2 cells were collected and resuspended in PBS and Matrigel (1:1) solution, and about 7.5×106 RL95-2 cells were subcutaneously inoculated on the right side of the back of BALB / c nude mice. When the average volume of the tumor reached about 100-150mm3, the animals were randomly divided into 3 groups according to the tumor size, and were divided into test compound RO7589831 treatment group, test compound 10 treatment group and vehicle control group, 6 in each group. Oral gavage was performed once a day, and a total of 20 days of administration was performed. Tumor volume and animal body weight were measured twice a week. The tumor volume (TV) was calculated according to the following formula: TV (tumor volume) = 1 / 2×a×b2, where a and b represent the length and width of the tumor, respectively.

[0726] The tumor growth curve is shown in Figure 1, and the body weight change of the experimental animals is shown in Figure 2. From the experimental results, it can be seen that the compound 10 of the present application has a significant inhibitory effect on the proliferation of RL95-2 human endometrial cancer cells in the subcutaneous xenotransplantation model, and the inhibitory effect on tumor growth is better than that of the current clinical compound RO7589831. No decrease in body weight of the test animals was found, indicating that the compound of the present application has good safety after administration.

Claims

1. A compound of Formula (I) or a stereoisomer, tautomer, deuterated isomer, or pharmaceutically acceptable salt thereof: ###0001### (I) ​ wherein: is selected from a single and a double bond as required to give each atom a normal valency; key denotes may exist as (Z)- or (E)-stereoisomers, wherein * indicates the point of attachment; ring A is a 5- to 12-membered heterocyclyl or a 5- to 6-membered heteroaryl, wherein said heterocyclyl or heteroaryl contains at least one nitrogen atom; X is selected from CR a and N; Z is selected from CR c and N; or = Z is absent, i.e. For Y is selected from CR b , N, NR d , S and O; Alternatively, Y and Z together form a C 3-7 Cycloalkyl, 3-7 membered heterocyclic or 5-6 membered heteroaryl, wherein C 3-7 Cycloalkyl, 3-7-membered heterocyclic, or 5-6-membered heteroaryl groups optionally further selected from one or more atoms selected from deuterium, hydroxyl, halogen, nitro, cyano, alkyl, haloalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, -OR 5 =O, -C(=O)R 5 -C(=O)OR 5 -OC(=O)R 5 -NR 6 R 7 -C(=O)NR 6 R 7 -S(=O)2NR 6 R 7 -N(R) 6 )C(=O)R 7 and -N(R) 6 )C(=O)OR 7 The substituents are replaced; R a , R b , R c , R d are each independently selected from the group consisting of a hydrogen atom, a halogen, a cyano group, a C 1-6 alkyl group and a C 3-6 cycloalkyl group, wherein said C 1-6 alkyl or C 3-6 cycloalkyl group is optionally further substituted by one or more substituents selected from the group consisting of a halogen, a hydroxy group, a cyano group and a C 1-6 alkoxy group; R 1 the same or different, each independently selected from the group consisting of a deuterium atom, a hydroxyl group, a halogen, a nitro group, a cyano group, a C 1-6 alkyl group, a C 2-6 alkenyl group, a C 2-6 alkynyl group, a C 3-8 cycloalkyl group, a 5-10 membered heterocyclyl group, a C 6-10 aryl group, a 5-6 membered heteroaryl group, -SF5, -OR 5 , -OC(=O)R 5 , -C(=O)R 5 , -C(=O)OR 5 , -N(R 6 )C(=O)R 7 , -N(R 6 )C(=O)OR 7 , -NR 6 R 7 , -C(=O)NR 6 R 7 , -S(=O) r NR 6 R 7 and -S(=O) r R 5 , wherein said C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3- 8cycloalkyl group, 5-10 membered heterocyclyl group, C 6-10 aryl group, 5-6 membered heteroaryl group is optionally further substituted by one or more substituents selected from the group consisting of a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, a haloalkyl group, a haloalkoxy group, a hydroxyalkyl group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, -OR 5 , =O, -C(=O)R 5 , -C(=O)OR 5 , -OC(=O)R 5 , -NR 6 R 7 , -C(=O)NR 6 R 7 , -S(=O)2NR 6 R 7 , -N(R 6 )C(=O)R 7 and -N(R 6 )C(=O)OR 7 ; and / or, two R 1 together with the same carbon atom to which they are attached form a -C(=0); R 2 is selected from the group consisting of hydrogen atom, deuterium atom, hydroxyl group, halogen, nitro group, cyano group, C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-8 cycloalkyl group, 5-10 membered heterocyclyl group, C 6-10 aryl group, 5-6 membered heteroaryl group, -SF5, -OR 5 , -OC(=O)R 5 , -C(=O)R 5 , -C(=O)OR 5 , -N(R 6 )C(=O)R 7 , -N(R 6 )C(=O)OR 7 , -NR 6 R 7 , -C(=O)NR 6 R 7 , -S(=O) r NR 6 R 7 and -S(=O) r R 5 , wherein said C 1-6 alkyl group, C 1-6 alkenyl group, C 1-6 alkynyl group, C 3-8 cycloalkyl group, 5-10 membered heterocyclyl group, C 6-10 aryl group, 5-6 membered heteroaryl group is optionally further substituted by one or more substituents selected from the group consisting of hydroxyl group, halogen, nitro group, cyano group, alkyl group, haloalkyl group, haloalkoxy group, hydroxyalkyl group, cycloalkyl group, heterocyclyl group, aryl group, heteroaryl group, -OR 5 , =O, -C(=O)R 5 , -C(=O)OR 5 , -OC(=O)R 5 , -NR 6 R 7 , -C(=O)NR 6 R 7 , -S(=O)2NR 6 R 7 , -N(R 6 )C(=O)R 7 and -N(R 6 )C(=O)OR 7 ; or R 2 and Y, R 2 and X are each independently taken together with the atom to which they are attached to form a C 3-7 cycloalkyl, 3-7 membered heterocyclyl, C 6-10 aryl or 5-6 membered heteroaryl, said C 3-7 cycloalkyl, 3-7 membered heterocyclyl, C 6-10 aryl or 5-6 membered heteroaryl is optionally further substituted with one or more substituents selected from a deuterium atom, hydroxy, halogen, nitro, cyano, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR 5 , =0, -C(=0)R 5 , -C(=0)OR 5 , -OC(=0)R 5 , -NR 6 R 7 , -C(=0)NR 6 R 7 , -S(=0)2NR 6 R 7 , -N(R 6 )C(=0)R 7 , and -N(R 6 )C(=0)OR 7 ; R g is selected from a hydrogen atom and a deuterium atom, preferably a hydrogen atom; R f each independently is selected from the group consisting of a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, and 5-10 membered heterocyclyl; wherein C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, and 5-10 membered heterocyclyl are optionally further substituted with one or more substituents selected from the group consisting of halogen, hydroxy, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and C 1-6 haloalkoxy; R 3a , R 3b are each independently selected from the group consisting of a hydrogen atom and a C 1-6 alkyl group; R 4 selected from C 1-6 alkyl, C 3-8 cycloalkyl and 5-10 membered heterocyclyl; wherein C 1-6 alkyl, C 3-8 cycloalkyl and 5-10 membered heterocyclyl are optionally further substituted by one or more substituents selected from halogen, hydroxy, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1- 6alkoxy and C 1-6 haloalkoxy; R 5 each independently is selected from the group consisting of an alkyl group, a cycloalkyl group, a heterocyclyl group, an aryl group, and a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclyl group, aryl group, and heteroaryl group are optionally further substituted with one or more substituents selected from the group consisting of a deuterium atom, a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a haloalkyl group, a haloalkoxy group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, =0, -C(=0)R 8 , -C(=0)0R 8 , -OC(=0)R 8 , -NR 9 R 10 , -C(=0)NR 9 R 10 , -S(=0)2NR 9 R 10 , and -N(R 9 )C(=0)R 10 ; R 6 and R 7 are each independently selected from the group consisting of a hydrogen atom, a hydroxyl group, an alkyl group, a cycloalkyl group, a heterocyclyl group, an aryl group and a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclyl group, aryl group and heteroaryl group are optionally further substituted with one or more substituents selected from the group consisting of a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, =0, -C(=0)R 8 , -C(=0)OR 8 , -OC(=0)R 8 , -NR 9 R 10 , -C(=0)NR 9 R 10 , -S(=0)2NR 9 R 10 and -N(R 9 )C(=0)R 10 ; or R 6 and R 7 together with the atom to which they are attached form a 4-8 membered heterocyclyl containing one or more N, O or S(=O) r wherein said 4-8 membered heterocyclyl is optionally further substituted by one or more substituents selected from the group consisting of hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -C(=O)R 8 , -C(=O)OR 8 , -OC(=O)R 8 , -NR 9 R 10 , -C(=O)NR 9 R 10 , -S(=O)2NR 9 R 10 and -N(R 9 )C(=O)R 10 ; R 8 , R 9 and R 10 are each independently selected from the group consisting of a hydrogen atom, an alkyl group, an amino group, a cycloalkyl group, a heterocyclyl group, an aryl group and a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclyl group, aryl group and heteroaryl group are optionally further substituted by one or more substituents selected from the group consisting of a hydroxyl group, a halogen, a nitro group, an amino group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, a carboxyl group and a carboxylate group; n is 0, 1, 2, 3, or 4; and each r is independently 0, 1, or 2.

2. The compound according to claim 1, or a stereoisomer, a tautomer, a deuterated isotope, or a pharmaceutically acceptable salt thereof, wherein, X is selected from CR a and N; Y is selected from CR b and N; Z is selected from CR c and N; and at least one of X, Y, Z is N and X, Y, Z are not simultaneously N; or Y and Z together form a C 3-7 Cycloalkyl, 3-7 membered heterocyclyl or 5-6 membered heteroaryl, said C 3-7 Cycloalkyl, 3-7 membered heterocyclyl or 5-6 membered heteroaryl, said C 5 , =0, -C(=0)R 5 , -C(=0)OR 5 , -OC(=0)R 5 , -NR 6 R 7 , -C(=0)NR 6 R 7 , -S(=0)2NR 6 R 7 , -N(R 6 )C(=0)R 7 , and -N(R 6 )C(=0)OR 7 ; R a , R b , R c , R 5 , R 6 and R 7 are as defined in claim 1.

3. The compound according to claim 1 or 2, or a stereoisomer, a tautomer, a deuterated isotope, or a pharmaceutically acceptable salt thereof, wherein Z is selected from CH.

4. The compound according to any one of claims 1-3, or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof, which is a compound according to Formula (II), or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof: ###00006### (II) wherein: X, Y are each independently selected from CH and N; and at least one of X, Y is N; is selected from a single and a double bond as required to give each atom a normal valency; R 3a , R 3b each independently is selected from a hydrogen atom; Rings A, R 1 R 2 R g R f R 4 and n are as defined in claim 1.

5. The compound according to any one of claims 1-4, or a stereoisomer, tautomer, deuterated product, or pharmaceutically acceptable salt thereof, wherein the compound is a stereoisomer, tautomer, deuterated product, or pharmaceutically acceptable salt thereof of general formula (III): wherein: X, Y are each independently selected from CH and N; and at least one of X, Y is N; is selected from a single and a double bond as required to give each atom a normal valency; R 3a , R 3b each independently is selected from a hydrogen atom; Rings A, R 1 R 2 R g R f R 4 and n are as defined in claim 1.

6. The compound according to claim 1 or 3, or a stereoisomer, a tautomer, a deuterated isotope, or a pharmaceutically acceptable salt thereof, wherein Z is CH, and X, Y are CH.

7. The compound according to claim 1, 3 or 6, or a stereoisomer, tautomer, deuterated product or pharmaceutically acceptable salt thereof, wherein the compound is a stereoisomer, tautomer, deuterated product or pharmaceutically acceptable salt thereof of general formula (IV): wherein: is selected from a single and a double bond as required to give each atom a normal valency; R 3a , R 3b each independently is selected from a hydrogen atom; Rings A, R 1 R 2 R g R f R 4 and n are as defined in claim 1.

8. The compound according to claim 7, or a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof, wherein the compound is a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof of general formula (IV-a): wherein: R 3a , R 3b , ring A, R 1 , R 2 , R g , R f , R 4 and n are as defined in claim 7.

9. The compound according to claim 1 or 2, or a stereoisomer, a tautomer, a deuterated isotope, or a pharmaceutically acceptable salt thereof, wherein Z is selected from N.

10. The compound of claim 1, 2, or 9, or a stereoisomer, tautomer, deuterated isomer, or pharmaceutically acceptable salt thereof, which is a compound of Formula (V) or a stereoisomer, tautomer, deuterated isomer, or pharmaceutically acceptable salt thereof: wherein, X, Y are each independently selected from CH and N; and X, Y are not simultaneously N; is selected from a single and a double bond as required to give each atom a normal valency; R 3a , R 3b each independently is selected from a hydrogen atom; Rings A, R 1 R 2 R g R f R 4 and n are as defined in claim 1.

11. The compound of claim 10, or a stereoisomer, a tautomer, a deuterated isotope, or a pharmaceutically acceptable salt thereof, which is a compound of Formula (V-a) or a stereoisomer, a tautomer, a deuterated isotope, or a pharmaceutically acceptable salt thereof: wherein, X, Y, R 3a , R 3b , ring A, R 1 , R 2 , R g , R f , R 4 and n are as defined in claim 10.

12. The compound according to claim 1, or a stereoisomer, a tautomer, a deuterated isotope, or a pharmaceutically acceptable salt thereof, wherein =Z is absent.

13. The compound of claim 1 or 12, or a stereoisomer, tautomer, deuterated isomer, or pharmaceutically acceptable salt thereof, which is a compound according to Formula (VI): ###00019### (VI) or a stereoisomer, tautomer, deuterated isomer, or pharmaceutically acceptable salt thereof. wherein: is selected from a single and a double bond as required to give each atom a normal valency; Y is selected from NR d , S and O; R d selected from the group consisting of a hydrogen atom and C 1-6 alkyl, wherein said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of halogen, hydroxy, cyano and C 1-6 alkoxy; R 2 selected from the group consisting of hydroxy, halogen, cyano, C 1-6 alkyl, C 3-8 cycloalkyl and C 1-6 alkoxy, wherein said C 1-6 alkyl, C 3-8 cycloalkyl and C 1-6 alkoxy are optionally further substituted by one or more halogen, C 1-6 alkyl or C 3-8 cycloalkyl. X is selected from CH and N; or R 2 and X together with the atom to which they are attached form C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 6-10 aryl or 5-6 membered heteroaryl are optionally further substituted with one or more substituents selected from the group consisting of deuterium atom, hydroxy, halogen, nitro, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and C 1-6 alkoxy; or R 2 and Y together with the atom to which it is attached forms a C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 6-10 aryl or 5-6 membered heteroaryl is optionally further substituted with one or more substituents selected from the group consisting of deuterium atom, hydroxy, halogen, nitro, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and C 1-6 alkoxy; Rings A, R 1 R 2 R g R f R 4 R 3a R 3b and n are as defined in claim 1.

14. The compound according to claim 13, which is a compound according to general formula (VI-a) or a stereoisomer, a tautomer, a deuterated analog, or a pharmaceutically acceptable salt thereof: ###00019### (VI-a) 14 wherein: Y, R 2 X, Y, ring A, R 1 R 2 R g R f R 4 R 3a R 3b and n are as defined in claim 13.

15. The compound according to claim 13, or a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof, wherein the compound is a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof of general formula (VI-1): wherein is selected from a single and a double bond as required to give each atom a normal valency; X1, X2, X3are each independently selected from CR e and N, and X1, X2, X3are not simultaneously N; Y is selected from NR d , S and O; R d selected from a hydrogen atom and C 1-6 alkyl; R e each independently is selected from the group consisting of a hydrogen atom, a deuterium atom, a hydroxyl group, a halogen, a nitro group, a cyano group, a C 1-6 alkyl group, a C 1-6 haloalkyl group, a C 3-8 cycloalkyl group and a C 1-6 alkoxy group; Rings A, R 1 R g R f R 4 R 3a R 3b and n are as defined in claim 1.

16. The compound according to claim 15, or a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof, wherein the compound is a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof of general formula (VI-1a): wherein, X1, X2, X3, Y, ring A, R 1 , R g , R f , R 4 , R 3a , R 3b and n are as defined in claim 15.

17. The compound according to claim 13, or a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof, wherein the compound is a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof of general formula (VI-2): wherein, X4, X5, X6are each independently selected from CR e and N, and X4, X5, X6are not simultaneously N; X is selected from CH and N; R e each independently is selected from the group consisting of a hydrogen atom, a deuterium atom, a hydroxyl group, a halogen, a nitro group, a cyano group, a C 1-6 alkyl group, a C 1-6 haloalkyl group, a C 3-8 cycloalkyl group and a C 1-6 alkoxy group; Rings A, R 1 R g R f R 4 R 3a R 3b and n are as defined in claim 1.

18. The compound according to claim 17, or a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof, wherein the compound is a stereoisomer, tautomer, deuterated derivative, or pharmaceutically acceptable salt thereof of general formula (VI-2a): X4, X5, X6, X, ring A, R 1 , R g , R f , R 4 , R 3a , R 3b and n are as defined in claim 17.

19. The compound of any one of claims 1-18, or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof, wherein is selected from the group consisting of:

20. The compound according to any one of claims 1-19, or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof, wherein R 1 each independently selected from C 6-10 aryl and C 3-8 cycloalkyl, wherein said C 6-10 aryl and C 3-8 cycloalkyl are optionally further substituted with one or more substituents selected from hydroxy, halo, cyano, amino, C 1-6 alkyl and C 1-6 alkoxy, and / or two R 1 with the same carbon atom to which they are attached forms a -C(=O).

21. The compound according to claim 20, or a stereoisomer, tautomer, deuterated analog, or pharmaceutically acceptable salt thereof, wherein R 1 each independently selected from C 6-10 aryl, more preferably phenyl, and / or two R 1 with the same carbon atom to which they are attached forms a -C(=O).

22. The compound of any one of claims 1-21, or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof, wherein is selected from the group consisting of: R 1a are each independently selected from the group consisting of C 6-10 aryl and C 3-8 cycloalkyl, wherein said C 6-10 aryl and C 3-8 cycloalkyl are optionally further substituted by one or more substituents selected from the group consisting of hydroxy, halogen, cyano, amino, C 1-6 alkyl and C 1-6 alkoxy, Preferably, R 1a selected from phenyl, More preferably, R 1a is phenyl.

23. The compound according to any one of claims 1-22, or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof, wherein R g is a hydrogen atom; R f is selected from the group consisting of C 1-6 alkyl and C 3-8 cycloalkyl, wherein said C 1-6 alkyl is optionally further substituted by one C 1-6 alkoxy, wherein said C 3-8 cycloalkyl is optionally further substituted by one or more halogen, and / or said C 3-8 cycloalkyl is preferably cyclopropyl or cyclobutyl.

24. The compound of any one of claims 1-23, or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof, wherein R 4 is selected from C 1-6 alkyl, preferably methyl.

25. The compound according to any one of claims 1-24, or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof, wherein R 2 is selected from the group consisting of a hydrogen atom, a hydroxyl group, a halogen, a cyano group, -NR 6 R 7 , a C 1-6 alkyl group, a C 3-8 cycloalkyl group and a C 1-6 alkoxy group, wherein said C 1-6 alkyl group, C 3-8 cycloalkyl group and C 1-6 alkoxy group is optionally further substituted with one or more halogen, C 1-6 alkyl group or C 3-8 cycloalkyl group; R 6 , R 7 are each independently selected from the group consisting of a hydrogen atom and a C 1-6 alkyl group.

26. The compound according to claim 25, or a stereoisomer, tautomer, deuterated analog, or pharmaceutically acceptable salt thereof, wherein R 2 is selected from a hydrogen atom, a hydroxyl group, a fluorine, a chlorine, a bromine, a cyano group, a methyl group, a tert-butyl group, a methoxy group, a trifluoromethyl group, a difluoromethyl group, a trifluoromethoxy group, a dimethylamino group, 27. The compound of any one of claims 1-26, or a stereoisomer, tautomer, deuterated isotope, or pharmaceutically acceptable salt thereof, wherein the compound is:

28. A pharmaceutical composition comprising a compound according to any one of claims 1-27, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

29. Use of a compound according to any one of claims 1-27, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 28, for the manufacture of a WRN inhibitor.

30. Use of a compound according to any one of claims 1-27, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 28, for the manufacture of a medicament for the treatment of a WRN-mediated disease; preferably wherein the WRN-mediated disease is a microsatellite instability-high cancer; further preferably the microsatellite instability-high cancer is selected from the group consisting of colorectal cancer, gastric cancer, endometrial cancer, rectal adenocarcinoma, adrenocortical carcinoma, uterine sarcoma, cervical cancer, nephroblastoma, mesothelioma, esophageal cancer, breast cancer, renal clear cell carcinoma, ovarian serous cystadenocarcinoma, cholangiocarcinoma, thymoma, liver cancer, head and neck squamous cell carcinoma, sarcoma, cutaneous melanoma, lung squamous cell carcinoma, prostate cancer, lung adenocarcinoma, bladder transitional cell carcinoma, pediatric neuroblastoma, chronic lymphocytic leukemia, and glioma, more preferably colorectal cancer, gastric cancer, or endometrial cancer.

31. Use of a compound according to any one of claims 1-27, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 28, for the manufacture of a medicament for the treatment of a microsatellite instability-high cancer; preferably wherein the microsatellite instability-high cancer is selected from the group consisting of colorectal cancer, gastric cancer, endometrial cancer, rectal adenocarcinoma, adrenocortical carcinoma, uterine sarcoma, cervical cancer, nephroblastoma, mesothelioma, esophageal cancer, breast cancer, renal clear cell carcinoma, ovarian serous cystadenocarcinoma, cholangiocarcinoma, thymoma, liver cancer, head and neck squamous cell carcinoma, sarcoma, cutaneous melanoma, lung squamous cell carcinoma, prostate cancer, lung adenocarcinoma, bladder transitional cell carcinoma, pediatric neuroblastoma, chronic lymphocytic leukemia, and glioma, further preferably colorectal cancer, gastric cancer, or endometrial cancer.

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